Pyrrolopyrazoles, potent kinase inhibitors

ABSTRACT

Pyrrole pyrazole compounds of formula I, compositions including these compounds and methods of their use are provided. Preferred compounds of formula I have activity as protein kinase inhibitors, including as inhibitors of PAK4.

This application is a Continuation of U.S. patent application Ser. No.11/813,658 filed on Mar. 17, 2008, which is a national stage filingunder 35 U.S.C. 371 of Patent Cooperation Treaty Patent Application No.PCT/IB2005/003975, filed Dec. 28, 2005, which claims the benefit of U.S.Provisional Patent Application Nos. 60/642,900 filed Jan. 10, 2005 and60/733,770 filed Nov. 4, 2005 the disclosures of which are incorporatedherein by reference in their entireties.

FIELD OF THE INVENTION

The present invention relates generally to novel chemical compounds andmethods. More particularly, the invention provides novel aminopyrrolopyrazole compounds and their analogs, having protein kinaseactivity, and methods of synthesizing and using such compounds.

BACKGROUND

Protein kinases are a family of enzymes that catalyze phosphorylation ofthe hydroxyl groups of specific tyrosine, serine, or threonine residuesin proteins. Typically, such phosphorylation can dramatically change thefunction of the protein and thus protein kinases can be pivotal in theregulation of a wide variety of cellular process, including metabolism,cell proliferation, cell differentiation, and cell survival. Themechanism of these cellular processes provides a basis for targetingprotein kinases to treat disease conditions resulting from or involvingdisorder of these cellular processes. Examples of such diseases include,but are not limited to, cancer and diabetes.

Protein kinases can be broken into two types, protein tyrosine kinases(PTKs) and serine-threonine kinases (STKs). Both PTKs and STKs can bereceptor protein kinases or non-receptor protein kinases. PAK is afamily of non-receptor STKs. The p21-activated protein kinase (PAK)family of serine/threonine protein kinases plays important roles incytoskeletal organization and cellular morphogenesis (Daniels et al.,Trends Biochem. Sci. 24: 350-355 (1999); Sells et al., Trends Cell.Biol. 7: 162-167 (1997)). PAK proteins were initially identified bytheir interaction with the active small GTPases, Cdc42, and Rac, andtheir homology to yeast kinase Step 20 (Manser et al., Nature 367: 40-46(1994)). In addition to mediating the regulation of actin cytoskeletonand cell adhesion by Cdc42 and Rac (Daniels et al., Trends Biochem. Sci.24: 350-355 (1999)), it was determined that some PAK proteins protectcells from apoptosis (Gnesutta et al., J. Biol. Chem. 276: 14414-14419(2001); Rudel et al., Science 276: 1571-1574 (1997); Schurmann et al.,Mol. Cell. Biol. 20: 453-461 (2000)); modulate mitogen activated protein(MAP) kinase pathways (Bagrodia et al., J. Biol. Chem. 270: 27995-27998(1995); Brown et al., Curr. Biol. 6: 598-605 (1996); Chaudhary et al.,Curr. Biol. 10: 551-554 (2000); Frost et al., EMBO J. 16: 6426-6438(1997); King et al., Nature 396: 180-183 (1998); Sun et al., Curr. Biol.10: 281-284 (2000)); mediate T-cell antigen receptor (TCR) signaling(Yablonski et al., EMBO J. 17: 5647-5657 (1998)); and respond to DNAdamage (Roig et al., J. Biol. Chem. 274: 31119-31122 (1999)). Throughthese diverse functions, PAK proteins regulate cell proliferation andmigration.

The full-length PAK4 nucleic acid and amino acid sequences are disclosedin U.S. Pat. No. 6,013,500 and have been deposited in GenBank underaccession numbers AF005046 (mRNA) and AAD01210 (amino acid). Modulationof human PAK4 activity is reported to result in alterations in cellularprocesses affecting cell growth and adhesion. For example,overexpression of PAK4 in fibroblasts leads to morphological changesthat are characteristic of oncogenic transformation through induction ofanchorage-independent growth and inhibition of apoptosis (Gnesutta etal., J. Biol. Chem. 276:14414-14419 (2001); Qu et al., Mol. Cell. Biol.21: 3523-2533 (2001)).

PAK4 is an attractive target for developing therapeutic agents effectivefor use in processes and disorders involving cytoskeletal alterations,such as, for example, cancer.

For other background references, see U.S. Patent Application PublicationNo. 2003/0171357 and PCT Publication WO02/12242.

SUMMARY

In one embodiment, the invention provides a compound of formula I,

wherein:

R¹ is chosen from —S(O)R^(a), —S(O)₂R^(a), C₁-C₁₂ alkyl, C₁-C₁₂ alkylsubstituted by 1 to 6 R⁵, C₃-C₁₂ cycloalkyl, C₃-C₁₂ cycloalkylsubstituted by 1 to 6 R⁵, C₂-C₁₂ alkenyl, C₂-C₁₂ alkenyl substituted by1 to 6 R⁵, C₄-C₁₂ cycloalkenyl, C₄-C₁₂ cycloalkenyl substituted by 1 to6 R⁵, C₂-C₁₂ alkynyl, C₂-C₁₂ alkynyl substituted by 1 to 6 R⁵, 3-12member heterocyclyl, 3-12 member heterocyclyl substituted by 1 to 6 R⁵,C₁-C₆ aralkyl, C₁-C₆ aralkyl substituted by 1 to 6 R⁵, C₁-C₆heteroaralkyl, C₁-C₆ heteroaralkyl substituted by 1 to 6 R⁵, C₆-C₁₀aryl, C₆-C₁₀ aryl substituted by 1 to 6 R⁵, 5-12 member heteroaryl, and5-12 member heteroaryl substituted by 1 to 6 R⁵, wherein any twoadjacent R⁵ together with the atoms to which they are attached may forma fused 4-7 member ring, and the said fused ring is optionally furthersubstituted by 1-3 R^(f);

R² and R³ are each independently chosen from —H, C₁-C₆ perfluoroalkyl,C₁-C₆ alkyl, C₃-C₆ cycloalkyl, —(C₁-C₃ alkylene)-(C₃-C₆ cycloalkyl),C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆ alkoxy, -(L)_(m)-halide,-(L)_(m)-CN, -(L)_(m)-OH, -(L)_(m)-NH₂, -(L)_(m)-(C₁-C₆ monoalkylamino)and -(L)_(m)-(C₂-C₈ dialkylamino), provided that R² and R³ are not bothH; or R² and R³ may form a ring selected from C₃-C₆ cycloalkyl, C₄-C₆cycloalkenyl and 3-6 member heterocyclyl, the said ring is optionallyfurther substituted by 1 to 2 groups selected from C₁-C₃ alkyl, C₁-C₃perfluoroalkyl, C₁-C₃ alkoxy, oxo, —(C₁-C₃ alkylene)_(m)-halide, —(C₁-C₃alkylene)_(m)-CN, —(C₁-C₃ alkylene)_(m)-OH, —(C₁-C₃ alkylene)_(m)-NH₂,—(C₁-C₃ alkylene)_(m)-(C₁-C₆ monoalkylamino) and —(C₁-C₃alkylene)_(m)-(C₂-C₈ dialkylamino);

R⁴ is selected from R^(a), —C(O)R^(a), —C(O)NR^(a)R^(b), —C(O)OR^(a),—C(O)CH(R^(t))R^(a), —C(O)NHCH(R^(a))R^(b), —C(O)OCH(R^(a))R^(b),—C(O)CH(R^(t))CH(R^(a))R^(b), —C(O)SR^(a), —S(O)R^(a), —S(O)NR^(a)R^(b),—S(O)OR^(a), —S(O)₂R^(a), —S(O)₂NR^(a)R^(b) and —S(O)₂OR^(a), whereinR^(t) is H or C₁-C₃ alkyl;

each R⁵ is independently selected from R^(c), -(L)_(m)-halide,-(L)_(m)-CN, -(L)_(m)-C(O)R^(c), -(L)_(m)-C(O)OR^(c),-(L)_(m)-C(O)NR^(c)R^(d), -(L)_(m)-C(O)SR^(c), -(L)_(m)-OR^(c),-(L)_(m)-OC(O)R^(c), -(L)_(m)-OC(O)NR^(c)R^(d), -(L)_(m)-O—C(O)OR^(c),-(L)_(m)-NO₂, -(L)_(m)-NR^(c)R^(d), -(L)_(m)-N(R^(c))C(O)R^(d),-(L)_(m)-N(R^(c))C(O)OR^(d), -(L)_(m)-NR^(c)S(O)R^(d),-(L)_(m)-NR^(c)S(O)OR^(d), -(L)_(m)-NR^(c)S(O)₂R^(d),-(L)_(m)-NR^(c)S(O)₂OR^(d), -(L)_(m)-SR^(c), -(L)_(m)-S(O)R^(c),-(L)_(m)-S(O)OR^(c), -(L)_(m)-S(O)₂R^(c), -(L)_(m)-S(O)₂OR^(c),-(L)_(m)-S(O)NR^(c)R^(d), -(L)_(m)-S(O)₂NR^(c)R^(d),-(L)_(m)-O-L-NR^(c)R^(d), -(L)_(m)-O-L-OR^(c) and-(L)_(m)-NR^(c)-L-OR^(d);

each R^(a), R^(b), R^(c), and R^(d) is independently selected from H,-(L)_(m)-(C₁-C₆ perfluoroalkyl), C₁-C₁₂alkyl, —(C₁-C₃alkylene)_(m)-(C₃-C₁₂ cycloalkyl), —(C₃-C₅ cycloalkylene)_(m)-(C₂-C₁₂alkenyl), -(L)_(m)-(C₄-C₁₂ cycloakenyl), —(C₃-C₅cycloalkylene)_(m)-(C₂-C₁₂alkynyl), -(L)_(m)-(3-12 member heterocyclyl),-(L)_(m)-(C₆-C₁₀ aryl) and -(L)_(m)-(5-12 member heteroaryl), eachR^(a), R^(b), R^(c) and R^(d) is independently optionally furthersubstituted by 1-6 R^(f); R^(a) and R^(b), or R^(c) and R^(d), togetherwith the atom to which they are attached, may optionally form a ringselected from 3-12 member heterocyclyl and 5-12 member heteroaryl, thesaid ring is optionally further substituted by 1-6 R^(f);

each R^(f) is independently selected from oxo, —(C₁-C₃alkylene)_(m)-(C₁-C₆ perfluoalkyl), C₁-C₁₂ alkyl, C₂-C₆ alkenyl, C₂-C₆alkynyl, —(C₁-C₃ alkylene)_(m)-(C₃-C₇ cycloalkyl), —(C₁-C₃alkylene)_(m)-(3-7 member heterocyclyl), —(C₁-C₃ alkylene)_(m)-(5-7member heteroaryl), -(L)_(m)-halide, -(L)_(m)-CN, -(L)_(m)-C(O)R^(k),-(L)_(m)-C(O)OR^(k), -(L)_(m)-C(O)NR^(k)R^(j), -(L)_(m)-OR^(k),-(L)_(m)-OC(O)R^(k), -(L)_(m)-NO₂, -(L)_(m)-NR^(k)R^(j),-(L)_(m)-N(R^(k))C(O)R^(j), -(L)_(m)-O-L-NR^(k)R^(j), -(L)_(m)-SR^(k),-(L)_(m)-S(O)R^(k), -(L)_(m)-S(O)₂R^(j)R^(k), each R^(f) isindependently optionally further substituted by 1-3 groups selected fromC₁-C₃ alkyl, halide and C₁-C₃ perfluoroalkyl;

each R^(k) and R^(j) is independently —H, —OH, C₁-C₃ perfluoroalkyl,C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₆ alkynyl, —(C₁-C₃ alkylene)_(m)-(C₃-C₆cycloalkyl) or —(C₁-C₃ alkylene)_(m)-(3 to 6 member heterocyclyl), R^(k)and R^(j) may optionally form a ring selected from 3-7 memberheterocyclyl and 5-7 member heteroaryl, the said ring is optionallyfurther substituted by 1 to 2 groups selected from C₁-C₃ alkyl, C₁-C₃perfluoroalkyl, C₁-C₃ alkoxy, oxo, —(C₁-C₃ alkylene)_(m)-halide, —(C₁-C₃alkylene)_(m)-CN, —(C₁-C₃ alkylene)_(m)-OH, —(C₁-C₃ alkylene)_(m)-NH₂,—(C₁-C₃ alkylene)_(m)-(C₁-C₆ monoalkylamino) and —(C₁-C₃alkylene)_(m)-(C₂-C₈ dialkylamino);

each L is independently a bivalent radical selected from —(C₁-C₆alkylene)-, —(C₃-C₇ cycloalkylene)-, —(C₁-C₆ alkylene)-(C₃-C₇cycloalkylene)- and —(C₃-C₇ cycloalkylene)-(C₁-C₆ alkylene)-;

each m is independently 0 or 1; and

n is 1, 2, or 3;

or a pharmaceutically acceptable salt, solvate or hydrate thereof.

In one particular aspect of the embodiment, and in combination of anyother particular aspect not inconsistent, n is 1. More particularly,each R² and R³ is independently selected from H, unsubstituted C₁-C₃alkyl and unsubstituted C₃-C₅ cycloalkyl, or R² and R³ form a ringselected from unsubstituted cyclopropyl, unsubstituted cyclobutyl andunsubstituted cyclopentyl. Even more particularly, R² is unsubstitutedmethyl, R³ is unsubstituted methyl.

In another particular aspect of the embodiment, and in combination ofany other particular aspect not inconsistent, R⁴ is selected from—C(O)NHCH(R^(a))R^(b), —C(O)OCH(R^(a))R^(b) and—C(O)CH(R^(t))CH(R^(a))R^(b). More particularly, R^(a) is selected from—(C₁-C₃ alkylene)_(m)-phenyl, —(C₁-C₃ alkylene)_(m)-(5-12 memberheteroaryl), —(C₁-C₃ alkylene)_(m)-(C₃-C₁₂ cycloalkyl) and —(C₁-C₃alkylene)_(m)-(3-12 member heterocyclyl), and R^(a) is optionallyfurther substituted by 1-6 R^(f); R^(b) is selected from C₁-C₆ alkylsubstituted by —NR^(j)R^(k), and —(C₁-C₃ alkylene)_(m)-(C₃-C₁₂heterocyclyl) optionally substituted by 1-6 R^(f). Even moreparticularly, R^(b) is a methyl group substituted by —NR^(j)R^(k). Yeteven more particularly, R^(a) is selected from phenyl, 5-12 memberheteroaryl, 3-12 member heterocyclyl and 3-12 member cycloalkyl, R^(a)is optionally further substituted by 1-6 R^(f), and R^(b) is a methylgroup substituted by NR^(j)R^(k).

In another particular aspect of the embodiment, and in combination ofany other particular aspect not inconsistent, R⁴ is selected from—C(O)NR^(a)R^(b), —C(O)OR^(a) and —C(O)CH(R)R^(a), wherein R^(b) isselected from H and C₁-C₃ alkyl, and R^(t) is selected from H and C₁-C₃alkyl. More particularly, R^(a) is selected from —(C₃-C₅cycloalkylene)-phenyl, —(C₃-C₅ cycloalkylene)-(5-12 member heteroaryl)and —(C₃-C₅ cycloalkylene)-(3-12 member heterocyclyl), and R^(a) isoptionally further substituted by 1-6 R^(f). Even more particularly,R^(a) is -(cyclopropylene)-phenyl, and R^(a) is optionally furthersubstituted by 1-6 R^(f).

In another particular aspect of the embodiment, and in combination ofany other particular aspect not inconsistent, n is 1, R⁴ is—C(O)NR^(a)R^(b), and wherein R^(a) and R^(b) form a ring selected from3-12 member heterocyclyl and 5-12 member heteroaryl, the said ringcontains 1-3 heteroatoms selected from N, O and S, and the said ring isoptionally further substituted by 1-6 R^(f). More particularly, the ringformed by R^(a) and R^(b) is selected from piperidinyl, morpholinyl,piperazinyl, pyridinyl and

and the ring is optionally further substituted by 1-6 R^(f).

In another particular aspect of the embodiment, and in combination ofany other particular aspect not inconsistent, R¹ is 5-12 memberheteroaryl, and R¹ is optionally further substituted by 1-6 R⁵. Moreparticularly, R¹ is selected from:

and R¹ is optionally further substituted as by 1-5 R⁵. Even moreparticularly, each R⁵ is independently -(L¹)_(m)-(C₁-C₆ perfluoalkyl),C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, —(C₁-C₃ alkylene)_(m)-(C₃-C₄cycloalkyl), —(C₁-C₃ alkylene)_(m)-(3-4 member heterocyclyl) optionallysubstituted by 1-2 C₁-C₃ alkyl, -(L¹)_(m)-halide, -(L¹)_(m)-CN,-(L¹)_(m)-C(O)R^(k), -(L¹)_(m)-C(O)OR^(k), -(L¹)_(m)-C(O)NR^(k)R^(j),-(L¹)_(m)-C(O)SR^(j), -(L¹)_(m)-OR^(k), -(L¹)_(m)-OC(O)R^(k),-(L¹)_(m)-OC(O)NR^(j)R^(k), -(L¹)_(m)-NO₂, -(L¹)_(m)-NR^(k)R^(j),N(R^(k))C(O)R^(j), -(L¹)_(m)-N(R^(k))C(O)OR^(j),-(L¹)_(m)-O-L¹-NR^(k)R^(j), -(L¹)_(m)-O-L¹-OR^(k),-(L¹)_(m)-NR^(j)-L¹-OR^(k), -(L¹)_(m)-SR^(k), -(L¹)_(m)-S(O)R^(k),-(L¹)_(m)-S(O)OR^(k), -(L¹)_(m)-S(O)NR^(j)R^(k), -(L¹)_(m)-S(O)₂R^(k),-(L¹)_(m)-S(O)₂OR^(k) or -(L¹)_(m)-S(O)₂NR^(j)R^(k), wherein each R^(j)and R^(k) is independently H, OH, C₁-C₃ alkyl or C₁-C₃ perfluoroalkyl,or R^(j) and R^(k) on the same nitrogen forms a 3-4 member ring selectedfrom aziridinyl and azetidinyl; L¹ is a bivalent radical selected from—(C₁-C₃ alkylene)-, —(C₃-C₄ cycloalkylene)-, -(3-4 memberheterocyclylene)-, —(C₁-C₃ alkylene)-(C₃-C₄ cycloalkylene)-, —(C₃-C₄cycloalkylene)-(C₁-C₃ alkylene)-, —(C₁-C₃ alkylene)-(3-4 memberheterocyclylene)- and -(3-4 member heterocyclylene)-(C₁-C₃ alkylene)-.Yet even more particularly, each R⁵ is independently halide or C₁-C₃alkyl.

In another embodiment, the present teachings provide a compound of theformula I,

wherein:

R¹ is chosen from —S(O)R^(a), —S(O)₂R^(a), C₁-C₁₂ alkyl, C₁-C₁₂ alkylsubstituted by at least one R^(f), C₃-C₁₂ cycloalkyl, C₃-C₁₂ cycloalkylsubstituted by at least one R^(f), C₂-C₁₂ alkenyl, C₂-C₁₂ alkenylsubstituted by at least one R^(f), C₂-C₁₂ alkynyl, C₂-C₁₂ alkynylsubstituted by at least one R^(f), 3-10 membered heterocycle, 3-10membered heterocycle substituted by at least one R^(f), C₁-C₆ aralkyl,C₁-C₆ aralkyl substituted by at least one R^(f), C₁-C₆ heteroaralkyl,C₁-C₆ heteroaralkyl substituted by at least one R^(f), C₆-C₁₀ aryl,C₆-C₁₀ aryl substituted by at least one R⁵, 5-10 membered heteroaryl,and 5-10 membered heteroaryl substituted by at least one R⁵, wherein anytwo adjacent R⁵ together with the atoms to which they are attached mayform a fused 4-7 membered ring;

R² and R³ are each independently chosen from —H, halide, —CN, —OH, —NO₂,—NH₂, C₁-C₃ perfluoroalkyl, C₁-C₃ alkoxy, C₁-C₆ alkoxyalkyl,unsubstituted C₁-C₆ aliphatic, C₁-C₆ alkylamine, C₁-C₁₂ alkyl, C₁-C₁₂alkyl substituted by 1 or 2 groups selected from —OH, —NH₂ and —CN,C₃-C₁₂ cycloalkyl, C₃-C₁₂ cycloalkyl substituted by 1 or 2 groupsselected from —OH, —NH₂ and —CN, C₂-C₁₂ alkenyl, C₂-C₁₂ alkenylsubstituted by 1 or 2 groups selected from —OH, —NH₂ and —CN, C₂-C₁₂alkynyl, and C₂-C₁₂ alkynyl substituted by 1 or 2 groups selected from—OH, —NH₂ and —CN;

R⁴ is selected from R^(a), —C(O)R^(a), —C(O)NHR^(a), —C(O)NR^(a)R^(b),—C(O)SR^(a), —S(O)R^(a), and —S(O)₂R^(a);

R⁵ is selected from R^(c), —OH, halide, —CN, —C(O)R^(c), —C(O)OR^(c),—C(O)NHR^(c), —C(O)NR^(c)R^(d), —OR^(c), —OC(O)R^(c), —NO₂,—NR^(c)R^(d), —N(R^(c))C(O)R^(d), —NHC(O)R^(c), —SR^(c), —S(O)R^(c), and—S(O)₂R^(c);

R^(a), R^(b), R^(c), and R^(d) are each independently selected fromC₁-C₁₂ alkyl, C₁-C₁₂ alkyl substituted by at least one R^(f), C₃-C₁₂cycloalkyl, C₃-C₁₂ cycloalkyl substituted by at least one R^(f), C₂-C₁₂alkenyl, C₂-C₁₂ alkenyl substituted by at least one R^(f), C₂-C₁₂alkynyl, C₂-C₁₂ alkynyl substituted by at least one R^(f), 3-10 memberedheterocycle, 3-10 membered heterocycle substituted by at least oneR^(f), C₆-C₁₀ aryl, C₆-C₁₀ aryl substituted by at least one R^(f), 5-10membered heteroaryl, 5-10 membered heteroaryl substituted by at leastone R^(f), C₁-C₆ aralkyl, C₁-C₆ aralkyl substituted by at least oneR^(f), C₁-C₆ heteroaralkyl, C₁-C₆ heteroaralkyl substituted by at leastone R^(f), and C₁-C₆ perfluoroalkyl; or R^(a) and R^(b), together withthe atom to which they are attached, form a 3 to 8 membered ring; orR^(c) and R^(d), together with the atom to which they are attached, forma 3 to 8 membered ring;

each R^(f), which may be the same or different, is selected from halide,—OH, —CN, —C(O)R^(k), —C(O)OR^(k), —C(O)NR^(k)R^(j), oxo, —OR^(k),—OC(O)R^(k), —NO₂, —NR^(k)R^(j), —N(R^(k))C(O)R^(j), —SR^(k),—S(O)R^(k), —S(O)₂R^(k), C₁-C₁₂ alkyl, C₁-C₁₂ alkyl substituted by atleast one R^(m), C₃-C₁₂ cycloalkyl, C₃-C₁₂ cycloalkyl substituted by atleast one R^(m), C₂-C₁₂ alkenyl, C₂-C₁₂ alkenyl substituted by at leastone R^(m), C₂-C₁₂ alkynyl, C₂-C₁₂ alkynyl substituted by at least oneR^(m), 3-10 membered heterocycle, 3-10 membered heterocycle substitutedby 1 to 4 R^(m), C₆-C₁₀ aryl, C₆-C₁₀ aryl substituted by 1 to 4 R^(m),5-10 membered heteroaryl, 5-10 membered heteroaryl substituted by 1 to 4R^(m), C₁-C₃ aralkyl, C₁-C₃ aralkyl substituted by 1 to 4 R^(m), C₁-C₃heteroaralkyl, C₁-C₃ heteroaralkyl substituted by 1 to 4 R^(m), andC₁-C₆ perfluoroalkyl;

R^(k) and R^(j) are each independently selected from —H, —OH, C₁-C₆aliphatic, and C₁-C₃ perfluoroalkyl; and

R^(m) is selected from halide, —OH, —CN, —C(O)R^(k), —C(O)OR^(k),—CONR^(k)R^(j), oxo, —OR^(k), —OC(O)R^(k), —NO₂, —NR^(k)R^(j),—N(R^(k))C(O)R^(k), —SR^(k), —S(O)R^(k), —S(O)₂R^(k), and C₁-C₃perfluoroalkyl;

n is 1, 2, or 3;

or a pharmaceutically acceptable salt, solvate or hydrate thereof.

In a particular aspect of this embodiment, n is 1.

In another embodiment, the present invention provides a compound offormula II,

wherein:

R² and R³ are each independently chosen from —H, halide, —CN, —OH, —NO₂,—NH₂, C₁-C₃ perfluoroalkyl, C₁-C₃ alkoxy, C₁-C₆ alkoxyalkyl,unsubstituted C₁-C₆ aliphatic, C₁-C₆ alkylamine, C₁-C₁₂ alkyl, C₁-C₁₂alkyl substituted by 1 or 2 groups selected from —OH, —NH₂ and —CN,C₃-C₁₂ cycloalkyl, C₃-C₁₂ cycloalkyl substituted by 1 or 2 groupsselected from —OH, —NH₂ and —CN, C₂-C₁₂ alkenyl, C₂-C₁₂ alkenylsubstituted by 1 or 2 groups selected from —OH, —NH₂ and —CN, C₂-C₁₂alkynyl, and C₂-C₁₂ alkynyl substituted by 1 or 2 groups selected from—OH, —NH₂ and —CN;

Ring A is selected from C₆-C₁₀ aryl, C₆-C₁₀ aryl substituted by at leastone R⁵, 5-10 membered heteroaryl, and 5-10 membered heteroarylsubstituted by at least one R⁵, wherein any two adjacent R⁵ togetherwith the atoms to which they are attached may form a fused 4-7 memberedring;

R⁵ is selected from R^(c), —OH, halide, —CN, —C(O)R^(c), —C(O)OR^(c),—C(O)NHR^(c), —C(O)NR^(c)R^(d), —OR^(c), —OC(O)R^(c), —NO₂, —NHR^(c),—NR^(c)R^(d), —N(R^(c))C(O)R^(d), —NHC(O)R^(c), —SR^(c), —S(O)R^(c), and—S(O)₂R^(c);

R^(a), R^(c), and R^(d) are each independently selected from C₁-C₁₂alkyl, C₁-C₁₂ alkyl substituted by at least one R^(f), C₃-C₁₂cycloalkyl, C₃-C₁₂ cycloalkyl substituted by at least one R^(f), C₂-C₁₂alkenyl, C₂-C₁₂ alkenyl substituted by at least one R^(f), C₂-C₁₂alkynyl, C₂-C₁₂ alkynyl substituted by at least one R^(f), 3-10 memberedheterocycle, 3-10 membered heterocycle substituted by at least oneR^(f), C₆-C₁₀ aryl, C₆-C₁₀ aryl substituted by at least one R^(f), 5-10membered heteroaryl, 5-10 membered heteroaryl substituted by at leastone R^(f), C₁-C₆ aralkyl, C₁-C₆ aralkyl substituted by at least oneR^(f), C₁-C₆ heteroaralkyl, C₁-C₆ heteroaralkyl substituted by at leastone R^(f), and C₁-C₆ perfluoroalkyl; or R^(c) and R^(d), together withthe atom to which they are attached, form a 3 to 8 membered ring;

each R^(f), which may be the same or different, is selected from halide,—OH, —CN, —C(O)R^(k), —C(O)OR^(k), —C(O)NR^(k)R^(j), oxo, —OR^(k),—OC(O)R^(k), —NO₂, —NR^(k)R^(j), —N(R^(k))C(O)R^(j), —SR^(k),—S(O)R^(k), —S(O)₂R^(k), C₁-C₁₂ alkyl, C₁-C₁₂ alkyl substituted by atleast one R^(m), C₃-C₁₂ cycloalkyl, C₃-C₁₂ cycloalkyl substituted by atleast one R^(m), C₂-C₁₂ alkenyl, C₂-C₁₂ alkenyl substituted by at leastone R^(m), C₂-C₁₂ alkynyl, C₂-C₁₂ alkynyl substituted by at least oneR^(m), 3-10 membered heterocycle, 3-10 membered heterocycle substitutedby 1 to 4 R^(m), C₆-C₁₀ aryl, C₆-C₁₀ aryl substituted by 1 to 4 R^(m),5-10 membered heteroaryl, 5-10 membered heteroaryl substituted by 1 to 4R^(m), C₁-C₃ aralkyl, C₁-C₃ aralkyl substituted by 1 to 4 R^(m), C₁-C₃heteroaralkyl, C₁-C₃ heteroaralkyl substituted by 1 to 4 R^(m), andC₁-C₆ perfluoroalkyl;

R^(k) and R^(j) are each independently selected from —H, —OH, C₁-C₆aliphatic, and C₁-C₃ perfluoroalkyl; and

R^(m) is selected from halide, —OH, —CN, —C(O)R^(k), —C(O)OR^(k),—CONR^(k)R^(j), oxo, —OR^(k), —OC(O)R^(k), —NO₂, —NR^(k)R^(j),—N(R^(k))C(O)R^(k), —SR^(k), —S(O)R^(k), —S(O)₂R^(k), and C₁-C₃perfluoroalkyl;

or a pharmaceutically acceptable salt, solvate or hydrate thereof.

In another embodiment, the present invention provides a compound offormula III,

wherein:

R² and R³ are each independently chosen from —H, halide, —CN, —OH, —NO₂,—NH₂, C₁-C₃ perfluoroalkyl, C₁-C₃ alkoxy, C₁-C₆ alkoxyalkyl,unsubstituted C₁-C₆ aliphatic, C₁-C₆ alkylamine, C₁-C₁₂ alkyl, C₁-C₁₂alkyl substituted by 1 or 2 groups selected from —OH, —NH₂ and —CN,C₃-C₁₂ cycloalkyl, C₃-C₁₂ cycloalkyl substituted by 1 or 2 groupsselected from —OH, —NH₂ and —CN, C₂-C₁₂ alkenyl, C₂-C₁₂ alkenylsubstituted by 1 or 2 groups selected from —OH, —NH₂ and —CN, C₂-C₁₂alkynyl, and C₂-C₁₂ alkynyl substituted by 1 or 2 groups selected from—OH, —NH₂ and —CN;

Ring A is selected from C₆-C₁₀ aryl, C₆-C₁₀ aryl substituted by at leastone R⁵, 5-10 membered heteroaryl, and 5-10 membered heteroarylsubstituted by at least one R⁵, wherein any two adjacent R⁵ togetherwith the atoms to which they are attached may form a fused 4-7 memberedring;

R⁵ is selected from R^(c), —OH, halide, —CN, —C(O)R^(c), —C(O)OR^(c),—C(O)NHR^(c), —C(O)NR^(c)R^(d), —OR^(c), —OC(O)R^(c), —NO₂, —NHR^(c),—NR^(c)R^(d), —N(R^(c))C(O)R^(d), —NHC(O)R^(c), —SR^(c), —S(O)R^(c), and—S(O)₂R^(c);

R^(a), R^(c), and R^(d) are each independently selected from C₁-C₁₂alkyl, C₁-C₁₂ alkyl substituted by at least one R^(f), C₃-C₁₂cycloalkyl, C₃-C₁₂ cycloalkyl substituted by at least one R^(f), C₂-C₁₂alkenyl, C₂-C₁₂ alkenyl substituted by at least one R^(f), C₂-C₁₂alkynyl, C₂-C₁₂ alkynyl substituted by at least one R^(f), 3-10 memberedheterocycle, 3-10 membered heterocycle substituted by at least oneR^(f), C₆-C₁₀ aryl, C₆-C₁₀ aryl substituted by at least one R^(f), 5-10membered heteroaryl, 5-10 membered heteroaryl substituted by at leastone R^(f), C₁-C₆ aralkyl, C₁-C₆ aralkyl substituted by at least oneR^(f), C₁-C₆ heteroaralkyl, C₁-C₆ heteroaralkyl substituted by at leastone R^(f), and C₁-C₆ perfluoroalkyl; or R^(c) and R^(d), together withthe atom to which they are attached, form a 3 to 8 membered ring;

each R^(f), which may be the same or different, is selected from halide,—OH, —CN, —C(O)R^(k), —C(O)OR^(k), —C(O)NR^(k)R^(j), oxo, —OR^(k),—OC(O)R^(k), —NO₂, —NR^(k)R^(j), —N(R^(k))C(O)R^(j), —SR^(k),—S(O)R^(k), —S(O)₂R^(k), C₁-C₁₂ alkyl, C₁-C₁₂ alkyl substituted by atleast one R^(m), C₃-C₁₂ cycloalkyl, C₃-C₁₂ cycloalkyl substituted by atleast one R^(m), C₂-C₁₂ alkenyl, C₂-C₁₂ alkenyl substituted by at leastone R^(m), C₂-C₁₂ alkynyl, C₂-C₁₂ alkynyl substituted by at least oneR^(m), 3-10 membered heterocycle, 3-10 membered heterocycle substitutedby 1 to 4 R^(m), C₆-C₁₀ aryl, C₆-C₁₀ aryl substituted by 1 to 4 R^(m),5-10 membered heteroaryl, 5-10 membered heteroaryl substituted by 1 to 4R^(m), C₁-C₃ aralkyl, C₁-C₃ aralkyl substituted by 1 to 4 R^(m), C₁-C₃heteroaralkyl, C₁-C₃ heteroaralkyl substituted by 1 to 4 R^(m), andC₁-C₆ perfluoroalkyl;

R^(k) and R^(j) are each independently selected from —H, —OH, C₁-C₆aliphatic, and C₁-C₃ perfluoroalkyl; and

R^(m) is selected from halide, —OH, —CN, —C(O)R^(k), —C(O)OR^(k),—CONR^(k)R^(j), oxo, —OR^(k), —OC(O)R^(k), —NO₂, —NR^(k)R^(j),—N(R^(k))C(O)R^(k), —SR^(k), —S(O)R^(k), —S(O)₂R^(k), and C₁-C₃perfluoroalkyl;

or a pharmaceutically acceptable salt, solvate or hydrate thereof.

In another embodiment, the present invention provides a compound offormula IV,

wherein:

R¹ is chosen from —S(O)R^(a), —S(O)₂R^(a), C₁-C₁₂ alkyl, C₁-C₁₂ alkylsubstituted by at least one R^(f), C₃-C₁₂ cycloalkyl, C₃-C₁₂ cycloalkylsubstituted by at least one R^(f), C₂-C₁₂ alkenyl, C₂-C₁₂ alkenylsubstituted by at least one R^(f), C₂-C₁₂ alkynyl, C₂-C₁₂ alkynylsubstituted by at least one R^(f), 3-10 membered heterocycle, 3-10membered heterocycle substituted by at least one R^(f), C₁-C₆ aralkyl,C₁-C₆ aralkyl substituted by at least one R^(f), C₁-C₆ heteroaralkyl,C₁-C₆ heteroaralkyl substituted by at least one R^(f), C₆-C₁₀ aryl,C₆-C₁₀ aryl substituted by at least one R⁵, 5-10 membered heteroaryl,and 5-10 membered heteroaryl substituted by at least one R⁵, wherein anytwo adjacent R⁵ together with the atoms to which they are attached mayform a fused 4-7 membered ring;

R² and R³ are each independently chosen from —H, halide, —CN, —OH, —NO₂,—NH₂, C₁-C₃ perfluoroalkyl, C₁-C₃ alkoxy, C₁-C₆ alkoxyalkyl,unsubstituted C₁-C₆ aliphatic, C₁-C₆ alkylamine, C₁-C₁₂ alkyl, C₁-C₁₂alkyl substituted by 1 or 2 groups selected from —OH, —NH₂ and —CN,C₃-C₁₂ cycloalkyl, C₃-C₁₂ cycloalkyl substituted by 1 or 2 groupsselected from —OH, —NH₂ and —CN, C₂-C₁₂ alkenyl, C₂-C₁₂ alkenylsubstituted by 1 or 2 groups selected from —OH, —NH₂ and —CN, C₂-C₁₂alkynyl, and C₂-C₁₂ alkynyl substituted by 1 or 2 groups selected from—OH, —NH₂ and —CN;

R⁵ is selected from R^(c), —OH, halide, —CN, —C(O)R^(c), —C(O)OR^(c),—C(O)NHR^(c), —C(O)NR^(c)R^(d), —OR^(c), —OC(O)R^(c), —NO₂, —NHR^(c),NR^(c)R^(d), —N(R^(c))C(O)R^(d), —NHC(O)R^(c), —SR^(c), —S(O)R^(c), and—S(O)₂R^(c);

R^(a), R^(c), and R^(d) are each independently selected from C₁-C₁₂alkyl, C₁-C₁₂ alkyl substituted by at least one R^(f), C₃-C₁₂cycloalkyl, C₃-C₁₂ cycloalkyl substituted by at least one R^(f), C₂-C₁₂alkenyl, C₂-C₁₂ alkenyl substituted by at least one R^(f), C₂-C₁₂alkynyl, C₂-C₁₂ alkynyl substituted by at least one R^(f), 3-10 memberedheterocycle, 3-10 membered heterocycle substituted by at least oneR^(f), C₆-C₁₀ aryl, C₆-C₁₀ aryl substituted by at least one R^(f), 5-10membered heteroaryl, 5-10 membered heteroaryl substituted by at leastone R^(f), C₁-C₆ aralkyl, C₁-C₆ aralkyl substituted by at least oneR^(f), C₁-C₆ heteroaralkyl, C₁-C₆ heteroaralkyl substituted by at leastone R^(f), and C₁-C₆ perfluoroalkyl; or R^(c) and R^(d), together withthe atom to which they are attached, form a 3 to 8 membered ring;

each R^(f), which may be the same or different, is selected from halide,—OH, —CN, —C(O)R^(k), —C(O)OR^(k), —C(O)NR^(k)R^(j), oxo, —OR^(k),—OC(O)R^(k), —NO₂, —NR^(k)R^(j), —N(R^(k))C(O)R^(j), —SR^(k),—S(O)R^(k), —S(O)₂R^(k), C₁-C₁₂ alkyl, C₁-C₁₂ alkyl substituted by atleast one R^(m), C₃-C₁₂ cycloalkyl, C₃-C₁₂ cycloalkyl substituted by atleast one R^(m), C₂-C₁₂ alkenyl, C₂-C₁₂ alkenyl substituted by at leastone R^(m), C₂-C₁₂ alkynyl, C₂-C₁₂ alkynyl substituted by at least oneR^(m), 3-10 membered heterocycle, 3-10 membered heterocycle substitutedby 1 to 4 R^(m), C₆-C₁₀ aryl, C₆-C₁₀ aryl substituted by 1 to 4 R^(m),5-10 membered heteroaryl, 5-10 membered heteroaryl substituted by 1 to 4R^(m), C₁-C₃ aralkyl, C₁-C₃ aralkyl substituted by 1 to 4 R^(m), C₁-C₃heteroaralkyl, C₁-C₃ heteroaralkyl substituted by 1 to 4 R^(m), andC₁-C₆ perfluoroalkyl;

R^(k) and R^(j) are each independently selected from —H, —OH, C₁-C₆aliphatic, and C₁-C₃ perfluoroalkyl;

R^(m) is selected from halide, —OH, —CN, —C(O)R^(k), —C(O)OR^(k),—CONR^(k)R^(j), oxo, —OR^(k), —OC(O)R^(k), —NO₂, —NR^(k)R^(j),—N(R^(k))C(O)R^(k), —SR^(k), —S(O)R^(k), —S(O)₂R^(k), and C₁-C₃perfluoroalkyl; and

when the atoms of any two adjacent R⁵ groups that are attached directlyto R¹ are chosen from carbon, nitrogen, oxygen and sulfur, then saidadjacent R⁵ groups may form a fused 4-7 membered ring;

or a pharmaceutically acceptable salt, solvate or hydrate thereof.

In another embodiment, the present invention provides a compound offormula V,

wherein:

R¹ is chosen from —S(O)R^(a), —S(O)₂R^(a), C₁-C₁₂ alkyl, C₁-C₁₂ alkylsubstituted by at least one R^(f), C₃-C₁₂ cycloalkyl, C₃-C₁₂ cycloalkylsubstituted by at least one R^(f), C₂-C₁₂ alkenyl, C₂-C₁₂ alkenylsubstituted by at least one R^(f), C₂-C₁₂ alkynyl, C₂-C₁₂ alkynylsubstituted by at least one R^(f), 3-10 membered heterocycle, 3-10membered heterocycle substituted by at least one R^(f), C₁-C₆ aralkyl,C₁-C₆ aralkyl substituted by at least one R^(f), C₁-C₆ heteroaralkyl,C₁-C₆ heteroaralkyl substituted by at least one R^(f), C₆-C₁₀ aryl,C₆-C₁₀ aryl substituted by at least one R⁵, 5-10 membered heteroaryl,and 5-10 membered heteroaryl substituted by at least one R⁵, wherein anytwo adjacent R⁵ together with the atoms to which they are attached mayform a fused 4-7 membered ring;

R² and R³ are each independently chosen from —H, halide, —CN, —OH, —NO₂,—NH₂, C₁-C₃ perfluoroalkyl, C₁-C₃ alkoxy, C₁-C₆ alkoxyalkyl,unsubstituted C₁-C₆ aliphatic, C₁-C₆ alkylamine, C₁-C₁₂ alkyl, C₁-C₁₂alkyl substituted by 1 or 2 groups selected from —OH, —NH₂ and —CN,C₃-C₁₂ cycloalkyl, C₃-C₁₂ cycloalkyl substituted by 1 or 2 groupsselected from —OH, —NH₂ and —CN, C₂-C₁₂ alkenyl, C₂-C₁₂ alkenylsubstituted by 1 or 2 groups selected from —OH, —NH₂ and —CN, C₂-C₁₂alkynyl, and C₂-C₁₂ alkynyl substituted by 1 or 2 groups selected from—OH, —NH₂ and —CN;

R⁵ is selected from R^(c), —OH, halide, —CN, —C(O)R^(c), —C(O)OR^(c),—C(O)NHR^(c), —C(O)NR^(c)R^(d), —OR^(c), —OC(O)R^(c), —NO₂,—NR^(c)R^(d), —N(R^(c))C(O)R^(d), —NHC(O)R^(c), —SR^(c), —S(O)R^(c), and—S(O)₂R^(c);

R^(a), R^(c), and R^(d) are each independently selected from C₁-C₁₂alkyl, C₁-C₁₂ alkyl substituted by at least one R^(f), C₃-C₁₂cycloalkyl, C₃-C₁₂ cycloalkyl substituted by at least one R^(f), C₂-C₁₂alkenyl, C₂-C₁₂ alkenyl substituted by at least one R^(f), C₂-C₁₂alkynyl, C₂-C₁₂ alkynyl substituted by at least one R^(f), 3-10 memberedheterocycle, 3-10 membered heterocycle substituted by at least oneR^(f), C₆-C₁₀ aryl, C₆-C₁₀ aryl substituted by at least one R^(f), 5-10membered heteroaryl, 5-10 membered heteroaryl substituted by at leastone R^(f), C₁-C₆ aralkyl, C₁-C₆ aralkyl substituted by at least oneR^(f), C₁-C₆ heteroaralkyl, C₁-C₆ heteroaralkyl substituted by at leastone R^(f), and C₁-C₆ perfluoroalkyl; or R^(c) and R^(d), together withthe atom to which they are attached, form a 3 to 8 membered ring;

each R^(f), which may be the same or different, is selected from halide,—OH, —CN, —C(O)R^(k), —C(O)OR^(k), —C(O)NR^(k)R^(j), oxo, —OR^(k),—OC(O)R^(k), —NO₂, —NR^(k)R^(j), —N(R^(k))C(O)R^(j), —SR^(k),—S(O)R^(k), —S(O)₂R^(k), C₁-C₁₂ alkyl, C₁-C₁₂ alkyl substituted by atleast one R^(m), C₃-C₁₂ cycloalkyl, C₃-C₁₂ cycloalkyl substituted by atleast one R^(m), C₂-C₁₂ alkenyl, C₂-C₁₂ alkenyl substituted by at leastone R^(m), C₂-C₁₂ alkynyl, C₂-C₁₂ alkynyl substituted by at least oneR^(m), 3-10 membered heterocycle, 3-10 membered heterocycle substitutedby 1 to 4 R^(m), C₆-C₁₀ aryl, C₆-C₁₀ aryl substituted by 1 to 4 R^(m),5-10 membered heteroaryl, 5-10 membered heteroaryl substituted by 1 to 4R^(m), C₁-C₃ aralkyl, C₁-C₃ aralkyl substituted by 1 to 4 R^(m), C₁-C₃heteroaralkyl, C₁-C₃ heteroaralkyl substituted by 1 to 4 R^(m), andC₁-C₆ perfluoroalkyl;

R^(k) and R^(j) are each independently selected from —H, —OH, C₁-C₆aliphatic, and C₁-C₃ perfluoroalkyl;

R^(m) is selected from halide, —OH, —CN, —C(O)R^(k), —C(O)OR^(k),—CONR^(k)R^(j), oxo, —OR^(k), —OC(O)R^(k), —NO₂, —NR^(k)R^(j),—N(R^(k))C(O)R^(k), —SR^(k), —S(O)R^(k), —S(O)₂R^(k), and C₁-C₃perfluoroalkyl; and

when the atoms of any two adjacent R⁵ groups that are attached directlyto R¹ are chosen from carbon, nitrogen, oxygen and sulfur, then saidadjacent R⁵ groups may form a fused 4-7 membered ring;

or a pharmaceutically acceptable salt, solvate or hydrate thereof.

In some embodiments, R² and R³ are —CH₃. In some embodiments, R⁴ isselected from —COR^(a), —C(O)NHR^(a), and —C(O)NR^(a)R^(b); and R^(a) isselected from C₁-C₁₂ alkyl, C₁-C₁₂ alkyl substituted by at least oneR^(f), C₃-C₁₂ cycloalkyl, C₃-C₁₂ cycloalkyl substituted by at least oneR^(f), C₂-C₁₂ alkenyl, C₂-C₁₂ alkenyl substituted by at least one R^(f),C₂-C₁₂ alkynyl, and C₂-C₁₂ alkynyl substituted by at least one R^(f). Insome embodiments, R^(a) is selected from cycloalkyl, and cycloalkylsubstituted by at least one R^(f). In some embodiments, R^(a) isselected from cyclopropyl, and cyclopropyl substituted by at least oneR^(f). In some embodiments, R^(a) is selected from cyclopropyl, andtrans-2-phenylcyclopropyl. In some embodiments, R^(a) is selected fromethyl and ethyl substituted by at least one R^(f). In some embodiments,R^(a) is

In some embodiments, R^(a) is

In some embodiments, R^(a) is selected from ethyl and ethyl substitutedby at least one R^(f). In some embodiments, R^(a) is

In some embodiments, R^(a) is

In some embodiments, R⁴ is

In some embodiments, R⁴ is

In some embodiments, R¹ is selected from 5-10 membered heteroaryl, and5-10 membered heteroaryl substituted by at least one R⁵. In someembodiments, Ring A is 5-10 membered heteroaryl. In some embodiments,Ring A is 5-10 membered heteroaryl substituted by at least one R⁵.

In some embodiments, the 5-10 membered heteroaryl is selected frompyrrole, furan, thiophene, oxazole, thiazole, pryazole, pyridine,pyrimidine, quinoline, isoquinoline, purine, tetrazole, triazine, andcarbazole.

In some embodiments, the 5-10 membered heteroaryl substituted by atleast one R⁵ is selected from

In some embodiments, the 5-10 membered heteroaryl substituted by atleast one R⁵ is selected from

In some embodiments, the 5-10 membered heteroaryl substituted by atleast one R⁵ is selected from

In some embodiments, the 5-10 membered heteroaryl substituted by atleast one R⁵ is selected from

In another embodiment, the current invention provides a compound offormula VI,

wherein:

B is a bond, —CHR^(t)—, —O— or —NH—, wherein R^(t) is H or C₁-C₃ alkyl;

R¹ is selected from

andR¹ is optionally further substituted by 1-5 R⁵;

R² is unsubstituted C₁-C₃ alkyl, R₃ is unsubstituted C₁-C₃ alkyl, or R²and R³ forms a ring selected from unsubstituted cyclopropyl andunsubstituted cyclobutyl;

each R⁵ is independently R^(x);

each R^(x) is independently -(L¹)_(m)-(C₁-C₆ perfluoalkyl), C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, —(C₁-C₃ alkylene)_(m)-(C₃-C₄ cycloalkyl),—(C₁-C₃ alkylene)_(m)-(3-4 member heterocyclyl) optionally substitutedby 1-2 C₁-C₃ alkyl, -(L¹)_(m)-halide, -(L¹)_(m)-CN, -(L¹)_(m)-C(O)R^(k),-(L¹)_(m)-C(O)OR^(k), -(L¹)_(m)-C(O)NR^(k)R^(j), -(L¹)_(m)-C(O)SR^(j),-(L¹)_(m)-OR^(k), -(L¹)_(m)-OC(O)R^(k), -(L¹)_(m)-OC(O)NR^(j)R^(k),-(L¹)_(m)-NO₂, -(L¹)_(m)-NR^(k)R^(j), -(L¹)_(m)-N(R^(k))C(O)R^(j),-(L¹)_(m)-N(R^(k))C(O)OR^(j), -(L¹)_(m)-O-L¹-NR^(k)R^(j),-(L¹)_(m)-O-L¹-OR^(k), -(L¹)_(m)-NR^(j)-L¹-OR^(k), -(L¹)_(m)-SR^(k),-(L¹)_(m)-S(O)R^(k), -(L¹)_(m)-S(O)OR^(k), -(L¹)_(m)-S(O)NR^(j)R^(k),-(L¹)_(m)-S(O)₂R^(k), -(L¹)_(m)-S(O)₂OR^(k) or-(L¹)_(m)-S(O)₂NR^(j)R^(k), wherein each R^(j) and R^(k) isindependently H, OH, C₁-C₃ alkyl or C₁-C₃ perfluoroalkyl, or R^(j) andR^(k) on the same nitrogen forms a 3-4 member ring selected fromaziridinyl and azetidinyl; L¹ is a bivalent radical selected from—(C₁-C₃ alkylene)-, —(C₃-C₄ cycloalkylene)-, -(3-4 memberheterocyclylene)-, —(C₁-C₃ alkylene)-(C₃-C₄ cycloalkylene)-, —(C₃-C₄cycloalkylene)-(C₁-C₃ alkylene)-, —(C₁-C₃ alkylene)-(3-4 memberheterocyclylene)- and -(3-4 member heterocyclylene)-(C₁-C₃alkylene)-;

R^(a) is selected from —(C₃-C₇ cycloalkylene)-phenyl, —(C₃-C₇cycloalkylene)-(5-12 member heteroaryl), —(C₃-C₇ cycloalkylene)-(3-12member heterocyclyl) and —(C₃-C₇ cycloalkyene)-(C₃-C₁₂ cycloalkyl),R^(a) is optionally further substituted by 1-6 groups selected from oxoand R^(x); and

each m is independently 0 or 1;

or a pharmaceutically acceptable salt, solvate or hydrate thereof.

In one particular aspect of this embodiment, and in combination of anyother particular aspect not inconsistent, B is —O—, R² is unsubstitutedmethyl, R³ is unsubstituted methyl. More particularly, R¹ is selectedfrom

Also more particularly, R¹ is selected from

In another particular aspect of the embodiment, and in combination ofany other particular aspect not inconsistent, B is —NH—, R² isunsubstituted methyl, R³ is unsubstituted methyl. More particularly, R¹is selected from

Also more particularly, R¹ is selected from

In another particular aspect of the embodiment, and in combination ofany other particular aspect not inconsistent, R^(a) is selected from-cyclopropylene-phenyl, -cyclopropylene-(5-12 member heteroaryl) and-cyclopropylene-(3-12 member heterocyclyl), R^(a) is optionally furthersubstituted by 1-6 groups selected from oxo and R^(x). Moreparticularly, R^(a) is selected from

wherein the stereochemistry indicated herein represents that the twosubstituents of the cyclopropylene group are trans, R^(a) is optionallyfurther substituted by 1-6 groups selected from oxo and R^(x). Even moreparticularly, the stereochemistry indicated herein represents theabsolute stereochemistry at the carbon centers of the cyclopropylenegroup.

In yet another embodiment, the current invention provides a compound offormula VII,

wherein:

B is a bond, —CHR^(t)—, —O— or —NH—, wherein R^(t) is H or C₁-C₃ alkyl;

R¹ is selected from

andR¹ is optionally further substituted by 1-5 R⁵;

R² is unsubstituted C₁-C₃ alkyl, R₃ is unsubstituted C₁-C₃ alkyl, or R²and R³ form a ring selected from unsubstituted cyclopropyl andunsubstituted cyclobutyl;

R^(a) is selected from -(L²)_(m)-phenyl, -(L²)_(m)-(5-12 memberheteroaryl), -(L²)_(m)-(C₃-C₁₂ cycloalkyl), and -(L²)_(m)-(3-12 memberheterocyclyl), wherein L² is a bivalent radical selected from —(C₁-C₃alkylene)-, —(C₃-C₄ cycloalkylene)-, —(C₁-C₃ alkylene)-(C₃-C₄cycloalkylene)-, —(C₃-C₄ cycloalkylene)-(C₁-C₃ alkylene)-, —O—, —(C₁-C₃alkylene)-O— and —O—(C₁-C₃ alkylene)-, and R^(a) is optionally furthersubstituted by 1-6 groups selected from oxo and R^(x);

R^(b) is —(C₁-C₆ alkylene)_(m)-NR^(p)R^(q), wherein each R^(p) and R^(q)is independently H, C₁-C₃ alkyl, or R^(p) and R^(q) forms a 3-7 memberheterocyclyl containing 1-2 heteroatoms selected from O and N, the said3-7 member heterocyclyl is optionally further substituted by 1-3 groupsselected from halide and C₁-C₃ alkyl;

each R⁵ is independently R^(x);

each R^(x) is independently -(L¹)_(m)-(C₁-C₆ perfluoalkyl), C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, —(C₁-C₃ alkylene)_(m)-(C₃-C₄ cycloalkyl),—(C₁-C₃ alkylene)_(m)-(3-4 member heterocyclyl) optionally substitutedby 1-2 C₁-C₃ alkyl, -(L¹)_(m)-halide, -(L¹)_(m)-CN, -(L¹)_(m)-C(O)R^(k),-(L¹)_(m)-C(O)OR^(k), -(L¹)_(m)-C(O)NR^(k)R^(j), -(L¹)_(m)-C(O)SR^(j),-(L¹)_(m)-OR^(k), -(L¹)_(m)-OC(O)R^(k), -(L¹)_(m)-OC(O)NR^(j)R^(k),-(L¹)_(m)-NO₂, -(L¹)_(m)-NR^(k)R^(j), -(L¹)_(m)-N(R^(k))C(O)R^(j),-(L¹)_(m)-N(R^(k))C(O)OR^(j), -(L¹)_(m)-O-L¹-NR^(k)R^(j),-(L¹)_(m)-O-L¹-OR^(k), -(L¹)_(m)-NR^(j)-L¹-OR^(k), -(L¹)_(m)-SR^(k),-(L¹)_(m)-S(O)R^(k), -(L¹)_(m)-S(O)OR^(k), -(L¹)_(m)-S(O)NR^(j)R^(k),-(L¹)_(m)-S(O)₂R^(k), -(L¹)_(m)-S(O)₂OR^(k) or-(L¹)_(m)-S(O)₂NR^(j)R^(k), wherein each R^(j) and R^(k) isindependently H, OH, C₁-C₃ alkyl or C₁-C₃ perfluoroalkyl, or R^(j) andR^(k) on the same nitrogen forms a 3-4 member ring selected fromaziridinyl and azetidinyl; L¹ is a bivalent radical selected from—(C₁-C₃ alkylene)-, —(C₃-C₄ cycloalkylene)-, -(3-4 memberheterocyclylene)-, —(C₁-C₃ alkylene)-(C₃-C₄ cycloalkylene)-, —(C₃-C₄cycloalkylene)-(C₁-C₃ alkylene)-, —(C₁-C₃ alkylene)-(3-4 memberheterocyclylene)- and -(3-4 member heterocyclylene)-(C₁-C₃alkylene)-;and

each m is independently 0 or 1;

or a pharmaceutically acceptable salt thereof.

In one particular aspect of the embodiment, and in combination of anyother particular aspect not inconsistent, B, R^(a), R^(b) and the carbonthat connects them form a S chiral center at the carbon. Moreparticularly, the compound is no less than 90% enantiomerically pureregarding the S chiral center.

In another particular aspect of the embodiment, and in combination ofany other particular aspect not inconsistent, B is —O—, R² isunsubstituted methyl, R³ is unsubstituted methyl. More particularly, R¹is selected from

Also more particularly, R¹ is selected from

In another particular aspect of the embodiment, and in combination ofany other particular aspect not inconsistent, B is —NH—, R² isunsubstituted methyl, R³ is unsubstituted methyl. More particularly, R¹is selected from

Also more particularly, R¹ is selected from

In another particular aspect of the embodiment, and in combination ofany other particular aspect not inconsistent, R^(b) is selected from—CH₂—N(CH₃)CH₃, —CH₂NHCH₃, —CH₂NH₂ and pyrrolyl.

In another particular aspect of the embodiment, and in combination ofany other particular aspect not inconsistent, R^(a) is selected fromphenyl, 5-12 member heteroaryl, 3-12 member heterocyclyl and 3-12 membercycloalkyl, R^(a) is optionally further substituted by 1-6 groupsselected from oxo and R^(x).

In yet another embodiment, the current invention provides apharmaceutical composition comprising a compound of the invention.

In yet another embodiment, the current invention provides apharmaceutical composition comprising a compound of the invention and apharmaceutically acceptable carrier.

In yet another embodiment, the current invention provides a method oftreating a mammalian disease condition mediated by protein kinaseactivity, comprising administering to a mammal a therapeuticallyacceptable amount of a compound, salt, hydrate or solvate of theinvention. In one aspect of this embodiment, mammalian disease conditionis tumor growth or abnormal cell proliferation.

In yet another embodiment, the current invention provides a method ofmodulating the activity of a protein kinase, comprising contacting theprotein kinase with an effective amount of a compound, orpharmaceutically acceptable salt, solvate of any of the invention. Inone aspect of this embodiment, the protein kinase is a PAK4 proteinkinase.

In some embodiments, the present teachings provide pharmaceuticalcompositions comprising any of the compounds described herein and apharmaceutically acceptable carrier. Examples of such compositions aredescribed below.

In some embodiments, the present teachings provide a method of treatingabnormal cell growth in a mammal, including a human, the methodcomprising administering to the mammal any of compound or pharmaceuticalcomposition of the present teachings.

In some embodiments, the abnormal cell growth is cancer, including, butnot limited to, lung cancer, bone cancer, pancreatic cancer, skincancer, cancer of the head or neck, cutaneous or intraocular melanoma,uterine cancer, ovarian cancer, rectal cancer, cancer of the analregion, stomach cancer, colon cancer, breast cancer, uterine cancer,carcinoma of the fallopian tubes, carcinoma of the endometrium,carcinoma of the cervix, carcinoma of the vagina, carcinoma of thevulva, Hodgkin's Disease, cancer of the esophagus, cancer of the smallintestine, cancer of the endocrine system, cancer of the thyroid gland,cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma ofsoft tissue, cancer of the urethra, cancer of the penis, prostatecancer, chronic or acute leukemia, lymphocytic lymphomas, cancer of thebladder, cancer of the kidney or ureter, renal cell carcinoma, carcinomaof the renal pelvis, neoplasms of the central nervous system (CNS),primary CNS lymphoma, spinal axis tumors, brain stem glioma, pituitaryadenoma, or a combination of one or more of the foregoing cancers. Insome embodiments, said abnormal cell growth is a benign proliferativedisease, including, but not limited to, psoriasis, benign prostatichypertrophy or restinosis.

In some embodiments, the method further comprises administering to themammal an amount of one or more substances selected from anti-tumoragents, anti-angiogenesis agents, signal transduction inhibitors, andantiproliferative agents, which amounts are together effective intreating said abnormal cell growth. Such substances include thosedisclosed in PCT Publication Nos. WO 00/38715, WO 00/38716, WO 00/38717,WO 00/38718, WO 00/38719, WO 00/38730, WO 00/38665, WO 00/37107 and WO00/38786, the disclosures of which are incorporated herein by referencein their entireties.

Examples of anti-tumor agents include mitotic inhibitors, for examplevinca alkaloid derivatives such as vinblastine vinorelbine, vindescineand vincristine; colchines allochochine, halichondrine,N-benzoyltrimethyl-methyl ether colchicinic acid, dolastatin 10,maystansine, rhizoxine, taxanes such as taxol (paclitaxel), docetaxel(Taxotere), 2′-N-[3-(dimethylamino)propyl]glutaramate (taxolderivative), thiocholchicine, trityl cysteine, teniposide, methotrexate,azathioprine, fluorouricil, cytocine arabinoside,2′2′-difluorodeoxycytidine (gemcitabine), adriamycin and mitamycin.Alkylating agents, for example cis-platin, carboplatin oxiplatin,iproplatin, Ethyl ester of N-acetyl-DL-sarcosyl-L-leucine (Asaley orAsalex), 1,4-cyclohexadiene-1,4-dicarbamic acid,2,5-bis(1-azirdinyl)-3,6-dioxo-, diethyl ester (diaziquone),1,4-bis(methanesulfonyloxy)butane (bisulfan or leucosulfan)chlorozotocin, clomesone, cyanomorpholinodoxorubicin, cyclodisone,dianhydroglactitol, fluorodopan, hepsulfam, mitomycin C,hycantheonemitomycin C, mitozolamide,1-(2-chloroethyl)-4-(3-chloropropyl)-piperazine dihydrochloride,piperazinedione, pipobroman, porfiromycin, spirohydantoin mustard,teroxirone, tetraplatin, thiotepa, triethylenemelamine, uracil nitrogenmustard, bis(3-mesyloxypropyl)amine hydrochloride, mitomycin,nitrosoureas agents such as cyclohexyl-chloroethylnitrosourea,methylcyclohexyl-chloroethylnitrosourea1-(2-chloroethyl)-3-(2,6-dioxo-3-piperidyl)-1-nitroso-urea,bis(2-chloroethyl)nitrosourea, procarbazine, dacarbazine, nitrogenmustard-related compounds such as mechloroethamine, cyclophosphamide,ifosamide, melphalan, chlorambucil, estramustine sodium phosphate,strptozoin, and temozolamide. DNA anti-metabolites, for example5-fluorouracil, cytosine arabinoside, hydroxyurea,2-[(3hydroxy-2-pyrinodinyl)methylene]-hydrazinecarbothioamide,deoxyfluorouridine, 5-hydroxy-2-formylpyridine thiosemicarbazone,alpha-2′-deoxy-6-thioguanosine, aphidicolin glycinate,5-azadeoxycytidine, beta-thioguanine deoxyriboside, cyclocytidine,guanazole, inosine glycodialdehyde, macbecin II, pyrazolimidazole,cladribine, pentostatin, thioguanine, mercaptopurine, bleomycin,2-chlorodeoxyadenosine, inhibitors of thymidylate synthase such asraltitrexed and pemetrexed disodium, clofarabine, floxuridine andfludarabine. DNA/RNA antimetabolites, for example, L-alanosine,5-azacytidine, acivicin, aminopterin and derivatives thereof such asN-[2-chloro-5-[[(2,4-diamino-5-methyl-6-quinazolinyl)-methyl]amino]benzoyl]-L-asparticacid,N-[4-[[(2,4-diamino-5-ethyl-6-quinazolinyl)methyl]amino]-benzoyl]-L-asparticacid,N-[2-chloro-4-[[(2,4-diaminopteridinyl)methyl]amino]benzoyl]-L-asparticacid, soluble Baker's antifol, dichloroallyl lawsone, brequinar, ftoraf,dihydro-5-azacytidine, methotrexate, N-(phosphonoacetyl)-L-aspartic acidtetrasodium salt, pyrazofuran, trimetrexate, plicamycin, actinomycin D,cryptophycin, and analogs such as cryptophycin-52 or, for example, oneof the preferred anti-metabolites disclosed in European PatentApplication No. 239362 such asN-(5-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]-2-thenoyl)-L-glutamicacid; growth factor inhibitors; cell cycle inhibitors; intercalatingantibiotics, for example adriamycin and bleomycin; proteins, for exampleinterferon; and anti-hormones, for example anti-estrogens such asNolvadex™ (tamoxifen) or, for example anti-androgens such as Casodex™(4′-cyano-3-(4-fluorophenylsulphonyl)-2-hydroxy-2-methyl-3′-(trifluoromethyl)propionanilide).Such conjoint treatment may be achieved by way of the simultaneous,sequential or separate dosing of the individual components of thetreatment.

Anti-angiogenesis agents include MMP-2 (matrix-metalloprotienase 2)inhibitors, MMP-9 (matrix-metalloprotienase 9) inhibitors, and COX-II(cyclooxygenase II) inhibitors. Examples of useful COX-II inhibitorsinclude CELEBREX™ (alecoxib), valdecoxib, and rofecoxib. Examples ofuseful matrix metalloproteinase inhibitors are described in WO 96/33172(published Oct. 24, 1996), WO 96/27583 (published Mar. 7, 1996),European Patent Application No. 97304971.1 (filed Jul. 8, 1997),European Patent Application No. 99308617.2 (filed Oct. 29, 1999), WO98/07697 (published Feb. 26, 1998), WO 98/03516 (published Jan. 29,1998), WO 98/34918 (published Aug. 13, 1998), WO 98/34915 (publishedAug. 13, 1998), WO 98/33768 (published Aug. 6, 1998), WO 98/30566(published Jul. 16, 1998), European Patent Publication 606,046(published Jul. 13, 1994), European Patent Publication 931,788(published Jul. 28, 1999), WO 90/05719 (published May 331, 1990), WO99/52910 (published Oct. 21, 1999), WO 99/52889 (published Oct. 21,1999), WO 99/29667 (published Jun. 17, 1999), PCT InternationalApplication No. PCT/IB98/01113 (filed Jul. 21, 1998), European PatentApplication No. 99302232.1 (filed Mar. 25, 1999), Great Britain patentapplication number 9912961.1 (filed Jun. 3, 1999), U.S. ProvisionalApplication No. 60/148,464 (filed Aug. 12, 1999), U.S. Pat. No.5,863,949 (issued Jan. 26, 1999), U.S. Pat. No. 5,861,510 (issued Jan.19, 1999), and European Patent Publication 780,386 (published Jun. 25,1997), all of which are herein incorporated by reference in theirentirety. Preferred MMP-2 and MMP-9 inhibitors are those that havelittle or no activity inhibiting MMP-1. More preferred, are those thatselectively inhibit MMP-2 and/or MMP-9 relative to the othermatrix-metalloproteinases MMP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7,MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13).

Examples of MMP inhibitors include AG-3340, RO 32-3555, RS 13-0830, andthe following compounds:3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-cyclopentyl)-amino]-propionicacid;3-exo-3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]octane-3-carboxylicacid hydroxyamide; (2R,3R)1-[4-(2-chloro-4-fluoro-benzyloxy)-benzenesulfonyl]-3-hydroxy-3-methyl-piperidine-2-carboxylicacid hydroxyamide;4-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-4-carboxylicacid hydroxyamide;3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-cyclobutyl)-amino]-propionicacid;4-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-4-carboxylicacid hydroxyamide;3-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-3-carboxylicacid hydroxyamide; (2R,3R)1-[4-(4-fluoro-2-methyl-benzyloxy)-benzenesulfonyl]-3-hydroxy-3-methyl-piperidine-2-carboxylicacid hydroxyamide;3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-1-methyl-ethyl)-amino]-propionicacid;3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(4-hydroxy-carbamoyltetrahydro-pyran-4-yl)-amino]-propionicacid;3-exo-3-[4-(4-chloro-phenoxy)benzene-sulfonylamino]-8-oxa-bicyclo[3.2.1]octane-3-carboxylicacid hydroxyamide;3-endo-3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]octane-3-carboxylicacid hydroxyamide;3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-tetrahydro-furan-3-carboxylicacid hydroxyamide; and pharmaceutically acceptable salts, solvates andhydrates thereof.

Examples of signal transduction inhibitors include agents that caninhibit EGFR (epidermal growth factor receptor) responses, such as EGFRantibodies, EGF antibodies, and molecules that are EGFR inhibitors; VEGF(vascular endothelial growth factor) inhibitors; and erbB2 receptorinhibitors, such as organic molecules or antibodies that bind to theerbB2 receptor, for example, HERCEPTIN™ (Genentech, Inc. of South SanFrancisco, Calif., USA).

EGFR inhibitors are described in, for example in WO 95/19970 (publishedJul. 27, 1995), WO 98/14451 (published Apr. 9, 1998), WO 98/02434(published Jan. 22, 1998), and U.S. Pat. No. 5,747,498 (issued May 5,1998). EGFR-inhibiting agents include, but are not limited to, themonoclonal antibodies C225 and anti-EGFR 22Mab (ImClone SystemsIncorporated of New York, N.Y., USA), the compounds ZD-1839(AstraZeneca), BIBX-1382 (Boehringer Ingelheim), MDX-447 (Medarex Inc.of Annandale, N.J., USA), and OLX-103 (Merck & Co. of WhitehouseStation, N.J., USA), VRCTC-310 (Ventech Research) and EGF fusion toxin(Seragen Inc. of Hopkinton, Mass.).

VEGF inhibitors, for example SU-5416 and SU-6668 (Sugen Inc. of SouthSan Francisco, Calif., USA), can also be combined or co-administeredwith the composition. VEGF inhibitors are described in, for example inWO 99/24440 (published May 20, 1999), PCT International ApplicationPCT/IB99/00797 (filed May 3, 1999), in WO 95/21613 (published Aug. 17,1995), WO 99/61422 (published Dec. 2, 1999), U.S. Pat. No. 5,834,504(issued Nov. 10, 1998), WO 98/50356 (published Nov. 12, 1998), U.S. Pat.No. 5,883,113 (issued Mar. 16, 1999), U.S. Pat. No. 5,886,020 (issuedMar. 23, 1999), U.S. Pat. No. 5,792,783 (issued Aug. 11, 1998), WO99/10349 (published Mar. 4, 1999), WO 97/32856 (published Sep. 12,1997), WO 97/22596 (published Jun. 26, 1997), WO 98/54093 (publishedDec. 3, 1998), WO 98/02438 (published Jan. 22, 1998), WO 99/16755(published Apr. 8, 1999), and WO 98/02437 (published Jan. 22, 1998), allof which are herein incorporated by reference in their entirety. Otherexamples of some specific VEGF inhibitors are IM862 (Cytran Inc. ofKirkland, Wash., USA); anti-VEGF monoclonal antibody bevacizumab(Genentech, Inc. of South San Francisco, Calif.); and angiozyme, asynthetic ribozyme from Ribozyme (Boulder, Colo.) and Chiron(Emeryville, Calif.).

ErbB2 receptor inhibitors, such as GW-282974 (Glaxo Wellcome plc), andthe monoclonal antibodies AR-209 (Aronex Pharmaceuticals Inc. of TheWoodlands, Tex., USA) and 2B-1 (Chiron), may be administered incombination with the composition. Such erbB2 inhibitors include thosedescribed in WO 98/02434 (published Jan. 22, 1998), WO 99/35146(published Jul. 15, 1999), WO 99/35132 (published Jul. 15, 1999), WO98/02437 (published Jan. 22, 1998), WO 97/13760 (published Apr. 17,1997), WO 95/19970 (published Jul. 27, 1995), U.S. Pat. No. 5,587,458(issued Dec. 24, 1996), and U.S. Pat. No. 5,877,305 (issued Mar. 2,1999), each of which is herein incorporated by reference in itsentirety. ErbB2 receptor inhibitors useful in the present invention arealso described in U.S. Provisional Application No. 60/117,341, filedJan. 27, 1999, and in U.S. Provisional Application No. 60/117,346, filedJan. 27, 1999, both of which are herein incorporated by reference intheir entirety.

Other antiproliferative agents that may be used include inhibitors ofthe enzyme farnesyl protein transferase and inhibitors of the receptortyrosine kinase PDGFr, including the compounds disclosed and claimed inthe following U.S. patent applications: Ser. No. 09/221,946 (filed Dec.28, 1998); Ser. No. 09/454,058 (filed Dec. 2, 1999); Ser. No. 09/501,163(filed Feb. 9, 2000); Ser. No. 09/539,930 (filed Mar. 31, 2000); Ser.No. 09/202,796 (filed May 22, 1997); Ser. No. 09/384,339 (filed Aug. 26,1999); and Ser. No. 09/383,755 (filed Aug. 26, 1999); and the compoundsdisclosed and claimed in the following U.S. provisional patentapplications: 60/168,207 (filed Nov. 30, 1999); 60/170,119 (filed Dec.10, 1999); 60/177,718 (filed Jan. 21, 2000); 60/168,217 (filed Nov. 30,1999), and 60/200,834 (filed May 1, 2000). Each of the foregoing patentapplications and provisional patent applications is herein incorporatedby reference in their entirety.

Compositions of the invention can also be used with other agents usefulin treating abnormal cell growth or cancer, including, but not limitedto, agents capable of enhancing antitumor immune responses, such asCTLA4 (cytotoxic lymphocite antigen 4) antibodies, and other agentscapable of blocking CTLA4; and anti-proliferative agents such as otherfarnesyl protein transferase inhibitors. Specific CTLA4 antibodies thatcan be used in the present invention include those described in U.S.Provisional Application 60/113,647 (filed Dec. 23, 1998) which is hereinincorporated by reference in its entirety.

Unless otherwise stated, the following terms used in the specificationand claims have the meanings discussed below. Variables defined in thissection, such as R, X, n and the like, are for reference within thissection only, and are not meant to have the save meaning as may be usedoutside of this definitions section. Further, many of the groups definedherein can be optionally substituted. The listing in this definitionssection of typical substituents is exemplary and is not intended tolimit the substituents defined elsewhere within this specification andclaims.

As used herein, the symbol [------] when incorporated into the chemicalstructure of a substituent means that the atom to which [------] isattached is the point of attachment of that substitutent to someposition on another molecule. For example, X in the hypotheticalmolecule CH₃CH₂—X might be defined as X is

In which case, the placement of [------] attached to the arbitrarilynumbered position C-1, means that C-1 of the phenyl ring is attached tothe methylene carbon.

The symbols “

” and “

”, when used together in a single molecule without further indicationotherwise, merely indicate relative stereochemistry of trans or ciswhere applicable. The symbol “

” and the symbol “

”, used together or separately, in combination with an indication ofthem representing the absolute stereochemistry, or an indication of “S”or “R” in the corresponding chemical structure or the accompanyingchemical name, indicate the absolute stereochemistry of thecorresponding chiral center.

“Aliphatic” refers to straight-chain, branched or cyclic C₁-C₁₂hydrocarbons which are completely saturated or which contains one ormore units of unsaturation but which are not aromatic. Examples ofaliphatic groups include linear, branched or cyclic alkyl, alkenyl,alkynyl groups and hybrids thereof such as (cycloalkyl)alkyl,(cycloalkenyl)alkyl, etc. An aliphatic group may be optionallysubstituted by 1-6 substituents. Suitable substituents on an aliphaticgroup include: 3-12 member heterocyclyl, C₆-C₁₀ aryl, 5-12 memberheteroaryl, halide, —NO₂, NH₂, NR₂, —CN, —COR, —COOR, —CONR₂, —OH, —OR,—OCOR, —SR, —SOR, —SO₂R, —SONR₂, —SO₂NR₂, wherein R is H, C₁-C₁₀ alkyl,3-10 member heterocyclyl, C₆-C₁₀ aryl, 5-12 member heteroaryl.

“C₁-C₁₂ alkyl” refers to a straight chain or branched saturatedhydrocarbon radical having from 1 to 12 carbon atoms. A C₁-C₁₂ alkylgroup may be optionally substituted by at least one substituent.Suitable substituents on a C₁-C₁₂ alkyl group include, but are notlimited to, 3-12 member heterocyclyl, C₆-C₁₀ aryl, 5-12 memberheteroaryl, halide, —NO₂, —NR₂, —CN, —COR, —COOR, —CONR₂, —OH, —OR,—OCOR, —SR, —SOR, —SO₂R, —SONR₂, —SO₂NR₂, wherein each R isindependently —H, C₁-C₁₀ alkyl, 3-12 member heterocyclyl, C₆-C₁₀ aryl,5-12 member heteroaryl. Examples of C₁-C₁₂ alkyl groups include, but arenot limited to methyl, ethyl, propyl, isopropyl, butyl, sec-butyl,iso-butyl, tert-butyl, pentyl, neo-pentyl, sec-pentyl, hexyl, heptyl,octyl, and the like, including substituted forms thereof. Further, theterm “alkyl” refers to a straight chain or branched saturatedhydrocarbon radical of 1 to 20 carbon atoms, or 1 to 12 carbon atoms, or1 to 8 carbon atoms, or 1 to 6 carbon atoms, or 1 to 4 carbon atoms.“Lower alkyl” refers specifically to an alkyl group having 1 to 4 carbonatoms. Alkyl may be substituted or unsubstituted. Suitable substituentson an alkyl group are the same as those described for a C₁-C₁₂ alkylgroup.

“Cycloalkyl” refers to a cyclic saturated hydrocarbon radical havingfrom 3 to 20 carbon atoms. A cycloalkyl group may be monocyclic andwhere permissible may be bicyclic or polycyclic. A cycloalkyl group maybe optionally substituted by at least one substituent. Suitablesubstituents on a cycloalkyl group are the same as those described foran alkyl group. Examples of cycloalkyl groups include, but are notlimited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, cyclooctyl, nobornyl, adamantyl, and the like, includingsubstituted forms thereof.

“Nonaromatic carbocyclyl” refers to a 3 to 12 member all-carbonmonocyclic ring group, all-carbon bicyclic or multicyclic ring systemgroup wherein one or more of the rings may contain one or more doublebonds but none of the rings has a completely conjugated pi-electronsystem. Examples, without limitation, of nonaromatic carbocyclyl arecyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl,cyclohexadienyl, adamantanyl, cycloheptyl, cycloheptatrienyl, and thelike. A nonaromatic carbocyclyl may be substituted or unsubstituted.Typical substituent groups are the same with those of alkyl group, asdefined herein. Illustrative examples of nonaromatic carbocyclyl arederived from, but not limited to, the following:

“Unsaturated nonaromatic carbocyclyl” refers to a nonaromaticcarbocyclyl, as defined herein, that contains at least one carbon carbondouble bond, one carbon carbon triple bond or a benzene ring.

“C₂-C₁₂ alkenyl” refers to a straight chain or branched unsaturatedhydrocarbon radical having from 2 to 12 carbon atoms. A C₂-C₁₂ alkenylgroup may have one or more points of unsaturation (i.e.—one or morecarbon-carbon double bonds). In the case where C₂-C₁₂ alkenyl has morethan one carbon-carbon double bond, the carbon-carbon double bonds canbe conjugated or unconjugated. A C₂-C₁₂ alkenyl group may be optionallysubstituted by at least one substituent. Suitable substituents on aC₂-C₁₂ alkenyl group are the same as those described for a C₁-C₁₂ alkylgroup. Examples of C₂-C₁₂ alkenyl include, but are not limited to,ethenyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, iso-butenyl, andthe like, including substituted forms thereof. Further, the term“alkenyl” refers to a straight chain or branched unsaturated hydrocarbonradical having from 2 to 20 carbon atoms, or 2 to 12 carbon atoms, or 2to 8 carbon atoms, or 2 to 6 carbon atoms, or 2 to 4 carbon atoms. Analkenyl group may have one or more points of unsaturation (i.e.—one ormore carbon-carbon double bonds). In the case where an alkenyl group hasmore than one carbon-carbon double bond, the carbon-carbon double bondscan be conjugated or unconjugated. An alkenyl group may be substitutedor unsubstituted. Suitable substituents on an alkenyl group are the sameas those described for a C₁-C₁₂ alkyl group.

“Alkoxy” refers to —OR^(c) wherein R^(c) is C₁-C₁₂ alkyl, C₂-C₁₂alkenyl, C₂-C₁₂ alkynyl, C₃-C₁₂ cycloalkyl or (C₁-C₆ alkylene)-(C₃-C₁₂cycloalkyl). A “C₁-C₁₂ alkoxy” refers to an alkoxy group, as definedherein, wherein R^(c) has 1 to 12 total carbon atoms.

“Alkoxyalkyl” refers to an alkyl, as defined herein, that is substitutedby at least one alkoxy group as defined herein. A “C₂-C₆ alkylalkoxy”refers an alkylalkoxy wherein the total carbon number of the alkyl andits alkoxy substituents are from 2 to 6.

“Alkylamino” refers to —NR^(p)R^(q) wherein each R^(p) and R^(q) isindependently H, C₁-C₁₂ alkyl, C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, C₃-C₁₂cycloalkyl, (C₁-C₆ alkylene)-(C₃-C₁₂ cycloalkyl) provided R^(p) andR^(q) are not both H. A “monoalkylamino” refers to an alkylamino group,as defined herein, wherein one of R^(p) and R^(q) is H. A “dialkylamino”refers to an alkylamino group, as defined herein, wherein none of R^(p)and R^(q) is H. A “C₁₋₁₂ alkylamino” refers to an alkylamino group thatcontains 1 to 10 carbon atoms.

“C₂-C₁₂ alkynyl” refers to a straight chain or branched hydrocarbonradical having from 2-12 carbon atoms and at least one carbon-carbontriple bond. In the case where C₂-C₁₂ alkynyl has more than onecarbon-carbon double bond, the carbon-carbon double bonds can beconjugated or unconjugated. A C₂-C₁₂ alkynyl group may be optionallysubstituted by at least one substituent. Suitable substituents on aC₂-C₁₂ alkynyl group are the same as those described for a C₁-C₁₂ alkylgroup. Examples of C₂-C₁₂ alkynyl include, but are not limited to,ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, and the like,including substituted forms thereof. Further, the term “alkynyl” refersto a straight chain or branched hydrocarbon radical of 2 to 20 carbonatoms, or 2 to 12 carbon atoms, or 2 to 8 carbon atoms, or 2 to 6 carbonatoms, or 2 to 4 carbon atoms, and having at least one carbon-carbontriple bond. Alkynyl may be substituted or unsubstituted. Suitablesubstituents on an alkynyl group are the same as those described for aC₁-C₁₂ alkyl group.

“Amino” refers to —NH₂.

“C₆-C₁₀ aryl” refers to an all-carbon monocyclic ring or polycyclic ringof 6 to 10 carbon atoms having a completely conjugated pi-electronsystem. A C₆-C₁₀ aryl group may be optionally substituted by at leastone substituent. Suitable substituents on a C₆-C₁₀ aryl group are thesame as those described for a C₁-C₁₂ alkyl group. Examples of C₆-C₁₀aryl include, but are not limited to, phenyl and naphthyl. Further, theterm “aryl” refers to an all-carbon monocyclic ring or polycyclic ringof 6 to 20 carbon atoms having a completely conjugated pi-electronsystem. The aryl group may be substituted or unsubstituted. Examples ofaryl include, but are not limited to, anthracenyl, phenanthreneyl andperylenyl.

“Aralkyl” refers to alkyl, as defined herein, that is substituted withan C₆₋₁₀ aryl group as defined above; e.g., —CH₂-phenyl, —(CH₂)₂-phenyl,—(CH₂)₃-phenyl, CH₃CH(CH₃)CH₂-phenyl, and the like and derivativesthereof. A C₁-C₆ aralkyl refers to a C₁-C₆ alkyl that is substitutedwith a C₆-C₁₀ aryl group.

“Heteroaralkyl” group means alkyl, as defined herein, that issubstituted with a 5-12 membered heteroaryl group; e.g., —CH₂pyridinyl,—(CH₂)₂pyrimidinyl, —(CH₂)₃imidazolyl, and the like, and derivativesthereof. A C₁-C₆ heteroaralkyl refers to a C₁-C₆ alkyl that issubstituted with an 5-12 membered heteroaryl group.

“Heteroaryl” refers to a monocyclic or fused ring group of 5 to 12 ringatoms containing one, two, three or four ring heteroatoms selected fromN, O, and S, the remaining ring atoms being C, and, in addition, havinga completely conjugated pi-electron system. Examples, withoutlimitation, of unsubstituted heteroaryl groups are pyrrole, furan,thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrimidine,quinoline, isoquinoline, purine, tetrazole, triazine, and carbazole. Theheteroaryl group may be substituted or unsubstituted. Typicalsubstituents include C₁₋₁₂ aliphatic, 3-10 membered heterocycle, 6-10membered aryl, halide, —NO₂, NH₂, NR₂, —CN, —COR, —COOR, —CONR₂, —OH,—OR, —OCOR, —SR, —SOR, —SO₂R, —SONR₂, —SO₂NR₂, wherein R is a C₁₋₁₀aliphatic, 3-10 membered heterocycle, C₆₋₁₀ aryl, 5-10 memberedheteroaryl.

A “pharmaceutically acceptable heteroaryl” is one that is sufficientlystable to be attached to a compound of the invention, formulated into apharmaceutical composition and subsequently administered to a patient inneed thereof.

Examples of typical monocyclic heteroaryl groups include, but are notlimited to:

Examples of bicyclic heteroaryl groups include, but are not limited to:

“Heteroalicyclic” or “heterocyclyl” refers to a monocyclic or polycyclicgroup having from 3 to 12 ring atoms, wherein from 1 to 4 ring atoms areheteroatoms selected from N, O, and S. “Heteroalicyclic” or“heterocyclyl” may also have one or more double bonds. However,“Heteroalicyclic” or “heterocyclyl” do not have a completely conjugatedpi-electron system. “Heteroalicyclic” or “heterocyclyl” can besubstituted or unsubstituted. Typical substituents include, but are notlimited to, C₁-C₁₂ aliphatic, 6-10 membered aryl, 6-10 membered aryl,halide, —NO₂, NH₂, NR₂, —CN, —COR, —COOR, —CONR₂, —OH, —OR, —OCOR, —SR,—SOR, —SO₂R, wherein R is a C₁-C₁₀ alkyl, 3-10 member heterocyclyl,C₆-C₁₀ aryl, 5-10 member heteroaryl.

Examples of saturated heterocyclyl groups include, but are not limitedto:

Examples of partially unsaturated heterocyclyl groups include, but arenot limited to:

When “ene” is added after “yl” at the end a term to form a new term, thenew term refers to a diradical formed by removing one hydrogen atom fromthe original term of which the new term derived from. For example, analkylene refers to a diradical group formed by removing one hydrogenatom from an alkyl group and that a “methylene” refers to a divalentradical —CH₂— derived from removing one hydrogen atom from methyl. Moreexamples of such diradicals include, but are not limited to: alkenylene,alkynylene, cycloalkylene, phenylene, heterocyclylene, heteroarylene and(nonaromatic unsaturated carbocyclylene), which are derived fromalkenyl, alkynyl, cycloalkyl, phenyl, heterocyclyl, heteroaryl and(nonaromatic unsaturated carbocyclyl), respectively. For example,“cyclopropylene” refers to both

For example, “C₁-C₃ alkylene” refers to all of the following: —CH₂—,—CH(CH₃)—, —CH₂—CH₂—, —CH₂—CH₂—CH₂—, —CH(CH₃)—CH₂— and —CH(CH₂CH₃)—.

“oxo” refers to an oxygen double bond “═O” substitution.

“Hydroxy” refers to —OH.

“Perfluoroalkyl” refers to an alkyl group in which all of its hydrogenatoms are replaced by fluorine atoms.

“Optional” or “optionally” means that the subsequently described eventor circumstance may but need not occur, and that the descriptionincludes instances where the event or circumstance occurs and instancesin which it does not. For example, “heterocycle group optionallysubstituted with an alkyl group” means that the alkyl may but need notbe present, and the description includes situations where theheterocycle group is substituted with an alkyl group and situationswhere the heterocycle group is not substituted with the alkyl group.

A “pharmaceutical composition” refers to a mixture of one or more of thecompounds described herein, or physiologically/pharmaceuticallyacceptable salts, solvates, hydrates or prodrugs thereof, with otherchemical components, such as physiologically/pharmaceutically acceptablecarriers and excipients. The purpose of a pharmaceutical composition isto facilitate administration of a compound to an organism.

As used herein, a “physiologically/pharmaceutically acceptable carrier”refers to a carrier or diluent that does not cause significantirritation to an organism and does not abrogate the biological activityand properties of the administered compound.

A “pharmaceutically acceptable excipient” refers to an inert substanceadded to a pharmaceutical composition to further facilitateadministration of a compound. Examples, without limitation, ofexcipients include calcium carbonate, calcium phosphate, various sugarsand types of starch, cellulose derivatives, gelatin, vegetable oils andpolyethylene glycols.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts that retain the biological effectiveness and properties ofthe parent compound. Such salts include:

(1) acid addition salts, which can be obtained by reaction of the freebase of the parent compound with inorganic acids such as hydrochloricacid, hydrobromic acid, nitric acid, phosphoric acid, sulfuric acid, andperchloric acid and the like, or with organic acids such as acetic acid,oxalic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid,ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaricacid, citric acid, succinic acid or malonic acid and the like; or

(2) salts formed when an acidic proton present in the parent compoundeither is replaced by a metal ion, e.g., an alkali metal ion, analkaline earth ion, or an aluminum ion; or coordinates with an organicbase such as ethanolamine, diethanolamine, triethanolamine,tromethamine, N-methylglucamine, and the like.

“PK” refers to receptor protein tyrosine kinase (RTKs), non-receptor or“cellular” tyrosine kinase (CTKs) and serine-threonine kinases (STKs).

“Modulation” or “modulating” refers to the alteration of the catalyticactivity of RTKs, CTKs and STKs. In particular, modulating refers to theactivation of the catalytic activity of RTKs, CTKs and STKs, preferablythe activation or inhibition of the catalytic activity of RTKs, CTKs andSTKs, depending on the concentration of the compound or salt to whichthe RTK, CTK or STK is exposed or, more preferably, the inhibition ofthe catalytic activity of RTKs, CTKs and STKs.

“Catalytic activity” refers to the rate of phosphorylation of tyrosineunder the influence, direct or indirect, of RTKs and/or CTKs or thephosphorylation of serine and threonine under the influence, direct orindirect, of STKs.

“Contacting” refers to bringing a compound of the present teachings anda target PK together in such a manner that the compound can affect thecatalytic activity of the PK, either directly, i.e., by interacting withthe kinase itself, or indirectly, i.e., by interacting with anothermolecule on which the catalytic activity of the kinase is dependent.Such “contacting” can be accomplished “in vitro,” i.e., in a test tube,a petri dish or the like. In a test tube, contacting may involve only acompound and a PK of interest or it may involve whole cells. Cells mayalso be maintained or grown in cell culture dishes and contacted with acompound in that environment. In this context, the ability of aparticular compound to affect a PK related disorder, i.e., the IC₅₀ ofthe compound, can be determined before use of compounds in vivo withmore complex living organisms is attempted. For cells outside theorganism, multiple methods exist, and are well-known to those skilled inthe art, to get the PKs in contact with the compounds including, but notlimited to, direct cell microinjection and numerous transmembranecarrier techniques.

“In vitro” refers to procedures performed in an artificial environmentsuch as, e.g., without limitation, in a test tube or culture medium.

“In vivo” refers to procedures performed within a living organism suchas, without limitation, a mouse, rat or rabbit.

“PK related disorder,” “PK driven disorder,” and “abnormal PK activity”all refer to a condition characterized by inappropriate, i.e., under or,more commonly, over, PK catalytic activity, where the particular PK canbe an RTK, a CTK or an STK. Inappropriate catalytic activity can ariseas the result of either: (1) PK expression in cells which normally donot express PKs, (2) increased PK expression leading to unwanted cellproliferation, differentiation and/or growth, or, (3) decreased PKexpression leading to unwanted reductions in cell proliferation,differentiation and/or growth. Over-activity of a PK refers to eitheramplification of the gene encoding a particular PK or production of alevel of PK activity which can correlate with a cell proliferation,differentiation and/or growth disorder (that is, as the level of the PKincreases, the severity of one or more of the symptoms of the cellulardisorder increases). Under-activity is, of course, the converse, whereinthe severity of one or more symptoms of a cellular disorder increase asthe level of the PK activity decreases.

“Treat”, “treating” and “treatment” refer to a method of alleviating orabrogating a PK mediated cellular disorder and/or its attendantsymptoms. With regard particularly to cancer, these terms simply meanthat the life expectancy of an individual affected with a cancer will beincreased or that one or more of the symptoms of the disease will bereduced.

“Organism” refers to any living entity comprised of at least one cell. Aliving organism can be as simple as, for example, a single eukarioticcell or as complex as a mammal, including a human being.

“Therapeutically effective amount” refers to that amount of the compoundbeing administered which will relieve to some extent one or more of thesymptoms of the disorder being treated. In reference to the treatment ofcancer, a therapeutically effective amount refers to that amount whichhas at least one of the following effects:

-   -   (1) reducing the size of the tumor;    -   (2) inhibiting (that is, slowing to some extent, preferably        stopping) tumor metastasis;    -   (3) inhibiting to some extent (that is, slowing to some extent,        preferably stopping) tumor growth, and    -   (4) relieving to some extent (or, preferably, eliminating) one        or more symptoms associated with the cancer.

“Monitoring” means observing or detecting the effect of contacting acompound with a cell expressing a particular PK. The observed ordetected effect can be a change in cell phenotype, in the catalyticactivity of a PK or a change in the interaction of a PK with a naturalbinding partner. Techniques for observing or detecting such effects arewell-known in the art. The effect is selected from a change or anabsence of change in a cell phenotype, a change or absence of change inthe catalytic activity of said protein kinase or a change or absence ofchange in the interaction of said protein kinase with a natural bindingpartner in a final aspect of this invention.

“Cell phenotype” refers to the outward appearance of a cell or tissue orthe biological function of the cell or tissue. Examples, withoutlimitation, of a cell phenotype are cell size, cell growth, cellproliferation, cell differentiation, cell survival, apoptosis, andnutrient uptake and use. Such phenotypic characteristics are measurableby techniques well-known in the art.

“Natural binding partner” refers to a polypeptide that binds to aparticular PK in a cell. Natural binding partners can play a role inpropagating a signal in a PK-mediated signal transduction process. Achange in the interaction of the natural binding partner with the PK canmanifest itself as an increased or decreased concentration of thePK/natural binding partner complex and, as a result, in an observablechange in the ability of the PK to mediate signal transduction.

DETAILED DESCRIPTION

Compounds of formulas I-VII can be made following the synthetic routesin Scheme 1 and Scheme 2. In Scheme 1 and Scheme 2 and the descriptionsfollowing, “BOC”, “Boc” or “boc” means N-tert-butoxycarbonyl, DCM meansCH₂Cl₂, DIPEA (also known as Hunig's base) means diisopropyl ethylamine, DMA means N,N-dimethylacetamide, “DMF” means dimethyl formamide,“DMSO” means dimethylsulfoxide, Et means —CH₂CH₃, “MTBE” means methylt-butyl ether, NMP means 1-methyl-2-pyrrolidinone, TEA means triethylamine, TFA means trifluoro acetic acid, THF means tetrahydrofuran. Whileschemes 1 and 2 and the description refer to compound I, schemes 1 and 2and the description are equally applicable to compounds II, III, IV, V,VI and VII.

Scheme 1 illustrates the synthesis of the intermediate I(H) used to makecompounds of formula I. The amino group of the substituted amino acidI(A) is alkylated to give compound I(B). This can typically be done bytreating compound I(A) with an alkylating agent in the presence of abase. An activated electrophilic double bond moiety is a commonly usedalkylating reagent. A typical reaction condition of alkylating I(A) withan activated electrophilic double bond moiety is to treat I(A) with theactivated double bond moiety in the presence of a strong base.Subsequent aqueous work up affords compound I(B). The amino group ofcompound I(B) is then protected with a boc group to give compound I(C).This can typically be done by treating compound I(B) with Boc agent inthe presence of a base. A typical condition is to treat compound I(B)with (Boc)₂O in the presence of Me₄NOH in MeCN as a solvent. Thecarboxylic acid group of compound I(C) is then converted into a methylester of compound I(D). A typical condition of converting the carboxylicacid group into the methyl ester group is to treat I(C) with methyliodide in DMF in the presence of a base. Compound I(D) then undergoes anintramolecular aldol condensation to give compound I(F). This cantypically be done by treating compound I(D) with a strong base in anaprotic solvent. A typical condition is to treat compound I(D) witht-BuOK in toluene. Subsequent aqueous workup gives compound I(F).Compound I(F) then undergoes a 2+3 cyclization with a hydrazine moietyto form compound I(G). A typical condition of the cyclization is toreflux compound I(F) with hydrazine and acetic acid in EtOH. The freebase pyrazole nitrogen of compound I(G) is then acylated to givecompound I(H). A typical condition of the acylation is to treat compoundI(G) with chloro ethyl carbonate in THF.

More detailed synthetic conditions of Scheme 1 can be found in U.S.Patent Application Publication No. 2003/0171357 and PCT Publication WO02/12242, the disclosure of which are incorporated herein by reference.

Scheme 2 illustrates two routes through which compounds of formula I canbe made from intermediate I(H). In the first route of Scheme 2, theethyl ester protecting group of the pyrazole nitrogen of I(H) is cleavedto give compound II(A). This reaction can typically be carried out bytreating the substrate I(H) with a base. A typical reaction condition isto reflux the substrate I(H) in dioxane and DCM in the presence of 2-3equivalents of LiOH followed by aqueous workup. Compound II(A) undergoesa nucleophilic reaction with an electrophilic R¹ moiety to give compoundII(B). This nucleophilic reaction can be alkylation, acylation,sulfonylation, reductive amination, and many other reactions that can becarried out for the pyrazole amino group of compound II(A). A typicalalkylation condition for the transformation of II(A) to II(B) is toreact II(A) with R¹—Cl in the presence of excess base, such as DMA andTEA at an elevated temperature of 80-140° C., and optionally undermicrowave radiation. Subsequent aqueous workup gives compound II(B). TheBoc group on the pyrrole nitrogen of compound II(B) is then removed togive compound II(C). This can typically be done by treating II(B) with astrong acid. A typical condition is to treat compound II(B) with 1:1TFA:DCM at room temperature for two hours. Subsequent aqueous work upaffords compound II(C). Alternatively, the transformation of compoundII(A) to compound II(C) can be carried out in a single step. Thealkylation of compound II(A) and the removal of the boc protecting groupof the pyrrole nitrogen can be carried out in an one pot reaction. Atypical reaction condition is to mix substrate II(A) with the alkylatingreagent R¹—Cl, excess DMA, one equivalent HCl, in dioxane at an elevatedtemperature and under microwave radiation. Compound II(C) then undergoesa nucleophilic reaction with an R⁴ electrophile to give compound I. Thenucleophilic reaction can be alkylation, acylation, sulfonylation,reductive amination and other reactions that a secondary alkyl amine cancarry out. An acylation reaction of compound II(C) can be carried out byreacting compound II(C) with an acylating R⁴ moiety. A typical acylationreaction condition is to react II(C) with an isocyanate R⁴ moiety in thepresence of TEA at room temperature. Subsequent aqueous workup givescompound I.

In the second route of Scheme 2, the boc group on the pyrrole nitrogenis removed to give compound III(A). The can typically be carried out bytreating compound I(H) with a strong acid. A typical reaction conditionis to treat compound I(H) with 4N HCl in dioxane and DCM. Subsequentaqueous workup affords compound III(A). Compound III(A) can thenundergoes a nucleophilic reaction with an R⁴ electrophile give compoundIII(B). Because the —NH₂ group attached to the pyrazole in compoundIII(A) is less reactive than the pyrrole nitrogen of III(A), thetransformation of III(A) to III(B) can be carried out without protectingthe pyrazole —NH₂ group of compound III(A). The nucleophilic reactioncarried out for this transformation can be an alkylation, acylation,sulfonylation, reductive amination. Relative mild reaction conditionsare preferred to achieve the reaction selectivity. An acylation reactionof III(A) to give III(B) is carried out by treating compound III(A) withan acylating reagent in the presence of base. A typical reactioncondition is to mix compound III(A) with excess of base, such as DIPEAin DCM and adding the resulting solution to an isocyanate at 0° C. Thereaction mixture is held at 0° C. for about two hours for the reactionto go complete. Subsequent aqueous workup gives compound III(B).

Selective acylation of the pyrrole nitrogen in the presence of theunprotected —NH₂ attached to the pyrazole to obtain compound III(B) canalso be done from compound I(H), through the intermediate of III(D). Thepyrrole nitrogen of I(H) is deprotected and further acylated to givecompound III(D). This can be done by treating compound I(H) with astrong acid, reducing the reaction mixture to a residue and thenreacting the residue with an acylating agent. A typical reactioncondition is to treat compound I(H) with 4N HCl in dioxane at roomtemperature for two hours and subsequently remove all solvent. Theresidue is dissolved and basified by a base such as DIPEA. The resultingsolution is then added to triphosgene at 0° C. The reaction mixture isallowed to stir at 0° C. for an hour. Subsequent aqueous work up affordscompound III(D). The crude compound III(D) then reacts with anucleophile to give compound III(B). A typical reaction condition is tomix compound III(D) with a R⁴ nucleophilic amine moiety in the presenceof a non-nucleophilic amine in DCM at room temperature. The reactionmixture is be held at room temperature for about two hours. Subsequentaqueous workup afford compound III(B).

The ethyl ester protecting group on the pyrazole nitrogen of compoundIII(B) is removed to give the free base compound III(C). This cantypically be done by treating compound III(B) with a base. A typicalreaction condition is to reflux compound III(B) in dioxane and DCM inthe presence of 2-3 equivalents of LiOH. Subsequent aqueous workupaffords compound III(C). Compound III(C) then undergoes a nucleophilicreaction with an R¹ electrophile moiety. This nucleophilic reaction canbe an acylation, alkylation, sulfonylation, reductive amination or oneof many other reactions that an amine functionality carries out. Atypical alkylation reaction condition is to treating compound III(C)with an alkylating agent such as R¹—Cl, in the presence of a base suchas 2 equivalents of DMA, in a solvent such as NMP, the reaction mixtureis then heated under microwave radiation to 140° C. for hour hours.Subsequent aqueous workup and purification gives compound of formula I.

Unless indicated otherwise, all references herein to the inventivecompounds include references to salts, solvates, hydrates and complexesthereof, and to solvates, hydrates and complexes of salts thereof,including polymorphs, stereoisomers, and isotopically labeled versionsthereof.

Pharmaceutically acceptable salts include acid addition and base salts(including disalts). Suitable acid addition salts are formed from acidswhich form non-toxic salts. Examples include the acetate, aspartate,benzoate, besylate, bicarbonate/carbonate, bisulphate/sulfate, borate,camsylate, citrate, edisylate, esylate, formate, fumarate, gluceptate,gluconate, glucuronate, hexafluorophosphate, hibenzate,hydrochloride/chloride, hydrobromide/bromide, hydroiodide/iodide,isethionate, lactate, malate, maleate, malonate, mesylate,methylsulfate, naphthylate, 2-napsylate, nicotinate, nitrate, orotate,oxalate, palmitate, pamoate, phosphate/hydrogen phosphate/dihydrogenphosphate, saccharate, stearate, succinate, tartrate, tosylate andtrifluoroacetate salts.

Suitable base salts are formed from bases which form non-toxic salts.Examples include the aluminum, arginine, benzathine, calcium, choline,diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine,potassium, sodium, tromethamine and zinc salts.

For a review on suitable salts, see “Handbook of Pharmaceutical Salts:Properties, Selection, and Use” by Stahl and Wermuth (Wiley-VCH,Weinheim, Germany, 2002), the disclosure of which is incorporated hereinby reference in its entirety.

A pharmaceutically acceptable salt of the inventive compounds can bereadily prepared by mixing together solutions of the compound and thedesired acid or base, as appropriate. The salt may precipitate fromsolution and be collected by filtration or may be recovered byevaporation of the solvent. The degree of ionization in the salt mayvary from completely ionized to almost non-ionized.

The compounds of the invention may exist in both unsolvated and solvatedforms. The term ‘solvate’ is used herein to describe a molecular complexcomprising the compound of the invention and one or morepharmaceutically acceptable solvent molecules, for example, ethanol. Theterm ‘hydrate’ is employed when the solvent is water. Pharmaceuticallyacceptable solvates in accordance with the invention include hydratesand solvates wherein the solvent of crystallization may be isotopicallysubstituted, e.g. D₂O, d₆-acetone, d₆-DMSO.

Also included within the scope of the invention are complexes such asclathrates, drug-host inclusion complexes wherein, in contrast to theaforementioned solvates, the drug and host are present in stoichiometricor non-stoichiometric amounts. Also included are complexes of the drugcontaining two or more organic and/or inorganic components which may bein stoichiometric or non-stoichiometric amounts. The resulting complexesmay be ionized, partially ionized, or non-ionized. For a review of suchcomplexes, see J Pharm Sci, 64 (8), 1269-1288 by Haleblian (August1975), the disclosure of which is incorporated herein by reference inits entirety.

Also within the scope of the invention are polymorphs, prodrugs, andisomers (including optical, geometric and tautomeric isomers) of theinventive compounds

Derivatives of compounds of the invention which may have little or nopharmacological activity themselves but can, when administered to apatient, be converted into the inventive compounds, for example, byhydrolytic cleavage. Such derivatives are referred to as ‘prodrugs’.Further information on the use of prodrugs may be found in ‘Pro-drugs asNovel Delivery Systems, Vol. 14, ACS Symposium Series (T Higuchi and WStella) and ‘Bioreversible Carriers in Drug Design’, Pergamon Press,1987 (ed. E B Roche, American Pharmaceutical Association), thedisclosures of which are incorporated herein by reference in theirentireties.

Prodrugs in accordance with the invention can, for example, be producedby replacing appropriate functionalities present in the inventivecompounds with certain moieties known to those skilled in the art as‘pro-moieties’ as described, for example, in “Design of Prodrugs” by HBundgaard (Elsevier, 1985), the disclosure of which is incorporatedherein by reference in its entirety.

Some examples of prodrugs in accordance with the invention include:

(i) where the compound contains a carboxylic acid functionality (—COOH),an ester thereof, for example, replacement of the hydrogen with(C₁-C₈)alkyl;

(ii) where the compound contains an alcohol functionality (—OH), anether thereof, for example, replacement of the hydrogen with(C₁-C₆)alkanoyloxymethyl; and

(iii) where the compound contains a primary or secondary aminofunctionality (—NH₂ or —NHR where R≠H), an amide thereof, for example,replacement of one or both hydrogens with (C₁-C₁₀)alkanoyl.

Further examples of replacement groups in accordance with the foregoingexamples and examples of other prodrug types may be found in theaforementioned references.

Finally, certain inventive compounds may themselves act as prodrugs ofother of the inventive compounds.

Compounds of the invention containing one or more asymmetric carbonatoms can exist as two or more stereoisomers. Where a compound of theinvention contains an alkenyl or alkenylene group, geometric cis/trans(or Z/E) isomers are possible. Where the compound contains, for example,a keto or oxime group or an aromatic moiety, tautomeric isomerism(‘tautomerism’) can occur. A single compound may exhibit more than onetype of isomerism.

Included within the scope of the invention are all stereoisomers,geometric isomers and tautomeric forms of the inventive compounds,including compounds exhibiting more than one type of isomerism, andmixtures of one or more thereof. Also included are acid addition or basesalts wherein the counterion is optically active, for example, D-lactateor L-lysine, or racemic, for example, DL-tartrate or DL-arginine.

Cis/trans isomers may be separated by conventional techniques well knownto those skilled in the art, for example, chromatography and fractionalcrystallization.

Conventional techniques for the preparation/isolation of individualenantiomers include chiral synthesis from a suitable optically pureprecursor or resolution of the racemate (or the racemate of a salt orderivative) using, for example, chiral high pressure liquidchromatography (HPLC).

Alternatively, the racemate (or a racemic precursor) may be reacted witha suitable optically active compound, for example, an alcohol, or, inthe case where the compound contains an acidic or basic moiety, an acidor base such as tartaric acid or 1-phenylethylamine. The resultingdiastereomeric mixture may be separated by chromatography and/orfractional crystallization and one or both of the diastereoisomersconverted to the corresponding pure enantiomer(s) by means well known toone skilled in the art.

Chiral compounds of the invention (and chiral precursors thereof) may beobtained in enantiomerically-enriched form using chromatography,typically HPLC, on an asymmetric resin with a mobile phase consisting ofa hydrocarbon, typically heptane or hexane, containing from 0 to 50%isopropanol, typically from 2 to 20%, and from 0 to 5% of an alkylamine,typically 0.1% diethylamine. Concentration of the eluate affords theenriched mixture.

Stereoisomeric conglomerates may be separated by conventional techniquesknown to those skilled in the art; see, for example, “Stereochemistry ofOrganic Compounds” by E L Eliel (Wiley, New York, 1994), the disclosureof which is incorporated herein by reference in its entirety.

The invention also includes isotopically-labeled compounds of theinvention, wherein one or more atoms is replaced by an atom having thesame atomic number, but an atomic mass or mass number different from theatomic mass or mass number usually found in nature. Examples of isotopessuitable for inclusion in the compounds of the invention includeisotopes of hydrogen, such as ²H and ³H, carbon, such as ¹¹C, ¹³C and¹⁴C, chlorine, such as ³⁶Cl, fluorine, such as ¹⁸F, iodine, such as ¹²³Iand ¹²⁵I, nitrogen, such as ¹³N and ¹⁵N, oxygen, such as ¹⁵O, ¹⁷O and¹⁸O, phosphorus, such as ³²P, and sulfur, such as ³⁵S. Certainisotopically-labeled compounds of the invention, for example, thoseincorporating a radioactive isotope, are useful in drug and/or substratetissue distribution studies. The radioactive isotopes tritium, ³H, andcarbon-14, ¹⁴C, are particularly useful for this purpose in view oftheir ease of incorporation and ready means of detection. Substitutionwith heavier isotopes such as deuterium, ²H, may afford certaintherapeutic advantages resulting from greater metabolic stability, forexample, increased in vivo half-life or reduced dosage requirements, andhence may be preferred in some circumstances. Substitution with positronemitting isotopes, such as ¹¹O, ¹⁸F, ¹⁵O and ¹³N, can be useful inPositron Emission Topography (PET) studies for examining substratereceptor occupancy.

Isotopically-labeled compounds of the invention can generally beprepared by conventional techniques known to those skilled in the art orby processes analogous to those described herein, using an appropriateisotopically-labeled reagent in place of the non-labeled reagentotherwise employed.

Pharmaceutically acceptable solvates in accordance with the inventioninclude those wherein the solvent of crystallization may be isotopicallysubstituted, e.g. D₂O, d₆-acetone, d₆-DMSO.

Compounds of the invention intended for pharmaceutical use may beadministered as crystalline or amorphous products, or mixtures thereof.They may be obtained, for example, as solid plugs, powders, or films bymethods such as precipitation, crystallization, freeze drying, spraydrying, or evaporative drying. Microwave or radio frequency drying maybe used for this purpose.

The compounds can be administered alone or in combination with one ormore other compounds of the invention, or in combination with one ormore other drugs (or as any combination thereof). Generally, they willbe administered as a formulation in association with one or morepharmaceutically acceptable excipients. The term “excipient” is usedherein to describe any ingredient other than the compound(s) of theinvention. The choice of excipient will to a large extent depend onfactors such as the particular mode of administration, the effect of theexcipient on solubility and stability, and the nature of the dosageform.

Pharmaceutical compositions suitable for the delivery of compounds ofthe invention and methods for their preparation will be readily apparentto those skilled in the art. Such compositions and methods for theirpreparation can be found, for example, in ‘Remington's PharmaceuticalSciences’, 19th Edition (Mack Publishing Company, 1995), the disclosureof which is incorporated herein by reference in its entirety.

Oral Administration

The compounds of the invention may be administered orally. Oraladministration may involve swallowing, so that the compound enters thegastrointestinal tract, or buccal or sublingual administration may beemployed by which the compound enters the blood stream directly from themouth.

Formulations suitable for oral administration include solid formulationssuch as tablets, capsules containing particulates, liquids, or powders,lozenges (including liquid-filled), chews, multi- and nano-particulates,gels, solid solution, liposome, films (including muco-adhesive), ovules,sprays and liquid formulations.

Liquid formulations include suspensions, solutions, syrups and elixirs.Such formulations may be used as fillers in soft or hard capsules andtypically include a carrier, for example, water, ethanol, polyethyleneglycol, propylene glycol, methylcellulose, or a suitable oil, and one ormore emulsifying agents and/or suspending agents. Liquid formulationsmay also be prepared by the reconstitution of a solid, for example, froma sachet.

The compounds of the invention may also be used in fast-dissolving,fast-disintegrating dosage forms such as those described in ExpertOpinion in Therapeutic Patents, 11 (6), 981-986 by Liang and Chen(2001), the disclosure of which is incorporated herein by reference inits entirety.

For tablet dosage forms, depending on dose, the drug may make up from 1wt % to 80 wt % of the dosage form, more typically from 5 wt % to 60 wt% of the dosage form. In addition to the drug, tablets generally containa disintegrant. Examples of disintegrants include sodium starchglycolate, sodium carboxymethyl cellulose, calcium carboxymethylcellulose, croscarmellose sodium, crospovidone, polyvinylpyrrolidone,methyl cellulose, microcrystalline cellulose, lower alkyl-substitutedhydroxypropyl cellulose, starch, pregelatinized starch and sodiumalginate. Generally, the disintegrant will comprise from 1 wt % to 25 wt%, preferably from 5 wt % to 20 wt % of the dosage form.

Binders are generally used to impart cohesive qualities to a tabletformulation. Suitable binders include microcrystalline cellulose,gelatin, sugars, polyethylene glycol, natural and synthetic gums,polyvinylpyrrolidone, pregelatinized starch, hydroxypropyl cellulose andhydroxypropyl methylcellulose. Tablets may also contain diluents, suchas lactose (monohydrate, spray-dried monohydrate, anhydrous and thelike), mannitol, xylitol, dextrose, sucrose, sorbitol, microcrystallinecellulose, starch and dibasic calcium phosphate dihydrate.

Tablets may also optionally include surface active agents, such assodium lauryl sulfate and polysorbate 80, and glidants such as silicondioxide and talc. When present, surface active agents are typically inamounts of from 0.2 wt % to 5 wt % of the tablet, and glidants typicallyfrom 0.2 wt % to 1 wt % of the tablet.

Tablets also generally contain lubricants such as magnesium stearate,calcium stearate, zinc stearate, sodium stearyl fumarate, and mixturesof magnesium stearate with sodium lauryl sulphate. Lubricants generallyare present in amounts from 0.25 wt % to 10 wt %, preferably from 0.5 wt% to 3 wt % of the tablet.

Other conventional ingredients include anti-oxidants, colorants,flavoring agents, preservatives and taste-masking agents.

Exemplary tablets contain up to about 80 wt % drug, from about 10 wt %to about 90 wt % binder, from about 0 wt % to about 85 wt % diluent,from about 2 wt % to about 10 wt % disintegrant, and from about 0.25 wt% to about 10 wt % lubricant.

Tablet blends may be compressed directly or by roller to form tablets.Tablet blends or portions of blends may alternatively be wet-, dry-, ormelt-granulated, melt congealed, or extruded before tabletting. Thefinal formulation may include one or more layers and may be coated oruncoated; or encapsulated.

The formulation of tablets is discussed in detail in “PharmaceuticalDosage Forms: Tablets, Vol. 1”, by H. Lieberman and L. Lachman, MarcelDekker, N.Y., N.Y., 1980 (ISBN 0-8247-6918-X), the disclosure of whichis incorporated herein by reference in its entirety.

Solid formulations for oral administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed-, sustained-, pulsed-, controlled-, targeted and programmedrelease.

Suitable modified release formulations are described in U.S. Pat. No.6,106,864. Details of other suitable release technologies such as highenergy dispersions and osmotic and coated particles can be found inVerma et al, Pharmaceutical Technology On-line, 25(2), 1-14 (2001). Theuse of chewing gum to achieve controlled release is described in WO00/35298. The disclosures of these references are incorporated herein byreference in their entireties.

Parenteral Administration

The compounds of the invention may also be administered directly intothe blood stream, into muscle, or into an internal organ. Suitable meansfor parenteral administration include intravenous, intraarterial,intraperitoneal, intrathecal, intraventricular, intraurethral,intrasternal, intracranial, intramuscular and subcutaneous. Suitabledevices for parenteral administration include needle (includingmicroneedle) injectors, needle-free injectors and infusion techniques.

Parenteral formulations are typically aqueous solutions which maycontain excipients such as salts, carbohydrates and buffering agents(preferably to a pH of from 3 to 9), but, for some applications, theymay be more suitably formulated as a sterile non-aqueous solution or asa dried form to be used in conjunction with a suitable vehicle such assterile, pyrogen-free water.

The preparation of parenteral formulations under sterile conditions, forexample, by lyophilization, may readily be accomplished using standardpharmaceutical techniques well known to those skilled in the art.

The solubility of compounds of the invention used in the preparation ofparenteral solutions may be increased by the use of appropriateformulation techniques, such as the incorporation ofsolubility-enhancing agents.

Formulations for parenteral administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed-, sustained-, pulsed-, controlled-, targeted and programmedrelease. Thus compounds of the invention may be formulated as a solid,semi-solid, or thixotropic liquid for administration as an implanteddepot providing modified release of the active compound. Examples ofsuch formulations include drug-coated stents and PGLA microspheres.

Topical Administration

The compounds of the invention may also be administered topically to theskin or mucosa, that is, dermally or transdermally. Typical formulationsfor this purpose include gels, hydrogels, lotions, solutions, creams,ointments, dusting powders, dressings, foams, films, skin patches,wafers, implants, sponges, fibers, bandages and microemulsions.Liposomes may also be used. Typical carriers include alcohol, water,mineral oil, liquid petrolatum, white petrolatum, glycerin, polyethyleneglycol and propylene glycol. Penetration enhancers may be incorporated;see, for example, J Pharm Sci, 88 (10), 955-958 by Finnin and Morgan(October 1999). Other means of topical administration include deliveryby electroporation, iontophoresis, phonophoresis, sonophoresis andmicroneedle or needle-free (e.g. Powderject™, Bioject™, etc.) injection.The disclosures of these references are incorporated herein by referencein their entireties.

Formulations for topical administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed-, sustained-, pulsed-, controlled-, targeted and programmedrelease.

Inhaled/Intranasal Administration

The compounds of the invention can also be administered intranasally orby inhalation, typically in the form of a dry powder (either alone, as amixture, for example, in a dry blend with lactose, or as a mixedcomponent particle, for example, mixed with phospholipids, such asphosphatidylcholine) from a dry powder inhaler or as an aerosol sprayfrom a pressurized container, pump, spray, atomizer (preferably anatomizer using electrohydrodynamics to produce a fine mist), ornebulizer, with or without the use of a suitable propellant, such as1,1,1,2-tetrafluoroethane or 1,1,1,2,3,3,3-heptafluoropropane. Forintranasal use, the powder may include a bioadhesive agent, for example,chitosan or cyclodextrin.

The pressurized container, pump, spray, atomizer, or nebulizer containsa solution or suspension of the compound(s) of the invention comprising,for example, ethanol, aqueous ethanol, or a suitable alternative agentfor dispersing, solubilizing, or extending release of the active, apropellant(s) as solvent and an optional surfactant, such as sorbitantrioleate, oleic acid, or an oligolactic acid.

Prior to use in a dry powder or suspension formulation, the drug productis micronized to a size suitable for delivery by inhalation (typicallyless than 5 microns). This may be achieved by any appropriatecomminuting method, such as spiral jet milling, fluid bed jet milling,supercritical fluid processing to form nanoparticles, high pressurehomogenisation, or spray drying.

Capsules (made, for example, from gelatin or HPMC), blisters andcartridges for use in an inhaler or insufflator may be formulated tocontain a powder mix of the compound of the invention, a suitable powderbase such as lactose or starch and a performance modifier such asl-leucine, mannitol, or magnesium stearate. The lactose may be anhydrousor in the form of the monohydrate, preferably the latter. Other suitableexcipients include dextran, glucose, maltose, sorbitol, xylitol,fructose, sucrose and trehalose.

A suitable solution formulation for use in an atomizer usingelectrohydrodynamics to produce a fine mist may contain from 1 μg to 20mg of the compound of the invention per actuation and the actuationvolume may vary from 1 μL to 100 μL. A typical formulation includes acompound of the invention, propylene glycol, sterile water, ethanol andsodium chloride. Alternative solvents which may be used instead ofpropylene glycol include glycerol and polyethylene glycol.

Suitable flavors, such as menthol and levomenthol, or sweeteners, suchas saccharin or saccharin sodium, may be added to those formulations ofthe invention intended for inhaled/intranasal administration.

Formulations for inhaled/intranasal administration may be formulated tobe immediate and/or modified release using, for example,poly(DL-lactic-coglycolic acid (PGLA). Modified release formulationsinclude delayed-, sustained-, pulsed-, controlled-, targeted andprogrammed release.

In the case of dry powder inhalers and aerosols, the dosage unit isdetermined by means of a valve which delivers a metered amount. Units inaccordance with the invention are typically arranged to administer ametered dose or “puff” containing a desired mount of the compound of theinvention. The overall daily dose may be administered in a single doseor, more usually, as divided doses throughout the day.

Rectal/Intravaginal Administration

Compounds of the invention may be administered rectally or vaginally,for example, in the form of a suppository, pessary, or enema. Cocoabutter is a traditional suppository base, but various alternatives maybe used as appropriate.

Formulations for rectal/vaginal administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed-, sustained-, pulsed-, controlled-, targeted and programmedrelease.

Ocular Administration

Compounds of the invention may also be administered directly to the eyeor ear, typically in the form of drops of a micronized suspension orsolution in isotonic, pH-adjusted, sterile saline. Other formulationssuitable for ocular and aural administration include ointments,biodegradable (e.g. absorbable gel sponges, collagen) andnon-biodegradable (e.g. silicone) implants, wafers, lenses andparticulate or vesicular systems, such as niosomes or liposomes. Apolymer such as crossed-linked polyacrylic acid, polyvinylalcohol,hyaluronic acid, a cellulosic polymer, for example,hydroxypropylmethylcellulose, hydroxyethylcellulose, or methylcellulose, or a heteropolysaccharide polymer, for example, gelan gum,may be incorporated together with a preservative, such as benzalkoniumchloride. Such formulations may also be delivered by iontophoresis.

Formulations for ocular/aural administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed-, sustained-, pulsed-, controlled-, targeted, or programmedrelease.

Other Technologies

Compounds of the invention may be combined with soluble macromolecularentities, such as cyclodextrin and suitable derivatives thereof orpolyethylene glycol-containing polymers, in order to improve theirsolubility, dissolution rate, taste-masking, bioavailability and/orstability for use in any of the aforementioned modes of administration.

Drug-cyclodextrin complexes, for example, are found to be generallyuseful for most dosage forms and administration routes. Both inclusionand non-inclusion complexes may be used. As an alternative to directcomplexation with the drug, the cyclodextrin may be used as an auxiliaryadditive, i.e. as a carrier, diluent, or solubilizer. Most commonly usedfor these purposes are alpha-, beta- and gamma-cyclodextrins, examplesof which may be found in PCT Publication Nos. WO 91/11172, WO 94/02518and WO 98/55148, the disclosures of which are incorporated herein byreference in their entireties.

Dosage

The amount of the active compound administered will be dependent on thesubject being treated, the severity of the disorder or condition, therate of administration, the disposition of the compound and thediscretion of the prescribing physician. However, an effective dosage istypically in the range of about 0.001 to about 100 mg per kg body weightper day, preferably about 0.01 to about 35 mg/kg/day, in single ordivided doses. For a 70 kg human, this would amount to about 0.07 toabout 7000 mg/day, preferably about 0.7 to about 2500 mg/day. In someinstances, dosage levels below the lower limit of the aforesaid rangemay be more than adequate, while in other cases still larger doses maybe used without causing any harmful side effect, with such larger dosestypically divided into several smaller doses for administrationthroughout the day.

Kit-of-Parts

Inasmuch as it may desirable to administer a combination of activecompounds, for example, for the purpose of treating a particular diseaseor condition, it is within the scope of the present invention that twoor more pharmaceutical compositions, at least one of which contains acompound in accordance with the invention, may conveniently be combinedin the form of a kit suitable for coadministration of the compositions.Thus the kit of the invention includes two or more separatepharmaceutical compositions, at least one of which contains a compoundof the invention, and means for separately retaining said compositions,such as a container, divided bottle, or divided foil packet. An exampleof such a kit is the familiar blister pack used for the packaging oftablets, capsules and the like.

The kit of the invention is particularly suitable for administeringdifferent dosage forms, for example, oral and parenteral, foradministering the separate compositions at different dosage intervals,or for titrating the separate compositions against one another. Toassist compliance, the kit typically includes directions foradministration and may be provided with a memory aid.

EXAMPLES

In the following examples and preparations, “BOC”, “Boc” or “boc” meansN-tert-butoxycarbonyl, DCM means CH₂Cl₂, DIPEA or DIEA means diisopropylethyl amine, DMA means N,N-dimethylacetamide, “DMF” means dimethylformamide, “DMSO” means dimethylsulfoxide, “DPPP” means1,3-bis(diphenylphosphino)propane, “HOAc” means acetic acid, “IPA” meansisopropyl alcohol. “MTBE” means methyl t-butyl ether, “NMP” means1-methyl 2-pyrrolidinone, TEA means triethyl amine, TFA means trifluoroacetic acid.

Specific Examples Example 16,6-dimethyl-N-[trans-2-phenylcyclopropyl]-3-(thieno[3,2-d]pyrimidin-4-ylamino)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide

Preparation of Compound 1a:N-(6,6-dimethyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)thieno[3,2-d]pyrimidin-4-amine

To a stirring solution of tert-butyl3-amino-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate(0.62 g, 2.46 mmol) in DMA (3 mL) was added4-chlorothieno[3,2-d]pyrimidine (0.44 g, 1.05 eq) and 4N HCl solution in1,4-dioxane (0.65 ml, 1.05 eq). The resulting mixture was heated to 140°C. for 0.5 hours in microwave reactor. It was cooled to room temperatureand the compound 1a was precipitated. Filtration and washing with CH₂Cl₂provided compound 1a as a yellow solid (0.48 g, 68% yield). Compound 1awas directly carried onto the next reaction without furtherpurification. LCMS (API-ES, M+H⁺): 287.0.

To a stirring mixture compound 1a (0.12 g, 0.42 mmol), and TEA (0.117ml, 2 eq) in DMSO (1 ml) and CH₂Cl₂ (2 ml) was addedtrans-2-phenylcyclopropyl isocyanate (0.068 ml, 1.1 eq). The resultingmixture was stirred at room temperature for 2 h. The reaction mixturewas purified by prep-HPLC to provide the title compound 1 as a whitesolid (0.019 g, 10%). ¹H NMR (CD₃OD) δ: 1.06 (m, 1H), 1.11 (m, 1H), 1.68(d, J=4.04 Hz, 6H), 1.98 (m, 1H), 2.69 (m, 1H), 4.43 (s, 2H), 7.01-7.07(m, 3H), 7.11-7.17 (m, 2H), 7.34 (d, J=5.56 Hz, 1H), 8.04 (d, J=5.31 Hz,1H), 8.59 (s, 1H). Anal. (C₂₃H₂₃N₇OS.0.3HOAc.0.8H₂O) C, H, N. HPLC: >95%purity.

Example 23-[(2-chlorothieno[3,2-d]pyrimidin-4-yl)amino]-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide

Preparation of Compound 2a:Tert-butyl-3-[(2-chlorothieno[3,2-d]pyrimidin-4-yl)amino]-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate

To a stirring solution of tert-butyl3-amino-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate(2.4 g, 9.5 mmol) in DMA (10 mL) was added2,4-dichlorothieno[3,2-d]pyrimidine (2.05 g, 1.05 eq) and triethylamine(2.64 ml, 2 eq). The resulting mixture was heated to 150° C. for 5minutes in microwave reactor. Saturated NaHCO₃ was added and the mixturewas extracted with ethyl acetate. The organic layer was dried oversodium sulfate, concentrated in vacuo. The residue was washed withmethylene chloride. Compound 2a (2.71 g, 68%) was obtained as a brownsolid and directly carried onto the next without further purification.LCMS (API-ES, M+H⁺): 421.

To a stirring mixture of compound 2a (0.102 g, 0.24 mmol) in CH₂Cl₂ (2ml), was added TFA (2 ml). The resulting mixture was stirred at roomtemperature for 2 h. After the reaction mixture was concentrated invacuo, a solution of TEA (135 ul, 4 eq) in MeCN (1 ml) CH₂Cl₂ (1 ml) wasadded and followed by trans-2-phenylcyclopropyl isocyanate. Theresulting mixture was stirred at room temperature for 1 h. The reactionmixture was purified by prep-HPLC to provide compound 2 as a white solid(0.021 g, 18%). ¹H NMR (CD₃OD) δ: 1.04-1.12 (m, 2H), 1.69 (d, J=3.28 Hz,2H), 1.95 (m, 1H), 2.67-2.73 (m, 1H), 4.48 (s, 2H), 7.01-7.06 (m, 3H),7.11-7.17 (m, 2H), 7.25 (d, J=5.31 Hz, 1H), 8.05 (d, J=4.55 Hz, 1H).Anal. (C₂₃H₂₂N₇OSCl.0.4HOAc.0.4H₂O) C, H, N. HPLC: >95% purity.

Example 3N-(5-{[(2S)-2-benzyl-4-methylpiperazin-1-yl]carbonyl}-6,6-dimethyl-1,4,5,6tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-2-chlorothieno[3,2-d]pyrimidin-4-amine

Preparation of 3a: ethyl3-amino-5-(chlorocarbonyl)-6,6-dimethyl-5,6-dihydropyrrolo[3,4-c]pyrazole-2(4H)-carboxylate

To a stirring mixture of 5-tert-butyl 2-ethyl3-amino-6,6-dimethylpyrrolo[3,4-c]pyrazole-2,5(4H,6H)-dicarboxylate(5.65 g, 17.4 mmol) in CH₂Cl₂ (20 ml) was added 4.0M HCl in dioxane (30ml). The resulting mixture was stirred at room temperature for 2 h.After the reaction mixture was concentrated in vacuo, a portion ofresidue (1.53 g, 5.15 mmol) was dissolved into a solution of DIPEA (3.6ml, 4 eq) in CH₂Cl₂ (10 ml). The resulting solution was slowly added toa solution of triphosgene (628 mg, 0.41 eq) in CH₂Cl₂ (10 ml) at 0° C.and the resulting mixture was stirred at 0° C. for 1 h. The reactionmixture was diluted with ethyl acetate and washed with saturated NaHCO₃,dried over sodium sulfate. The organic layer was filtered and evaporatedin vacuo to give a residue, compound 3a. Compound 3a was directlycarried onto the next reaction without further purification.

Preparation of 3b:5-{[(2S)-2-benzyl-4-methylpiperazin-1-yl]carbonyl}-6,6-dimethyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-amine

A portion of above residue (525 mg) was added into a solution of(3S)-3-benzyl-1-methylpiperazine (552 mg, 1.5 eq) in CH₂Cl₂ (4 ml). Theresulting mixture was stirred at reflux for 4 h. The reaction mixturewas then cooled to room temperature, and the solvent was removed invaccuo. To the residue was added 2N LiOH (3 ml) and methanol (2 ml). Theresulting mixture was stirred at reflux for 4 hours. The reactionmixture was purified by prep-HPLC to provide compound 3b as a whitesolid (150 mg). To a stirring solution of compound 3b (0.15, 0.41 mmol)in NMP (1 mL) was added 2,4-dichlorothieno[3,2-d]pyrimidine (0.084 g, 1eq) and triethylamine (0.11 ml, 2 eq). The resulting mixture was heatedto 140° C. for 5 minutes in microwave reactor. The reaction mixture waspurified by prep-HPLC to provide compound 3 as a white solid (0.014 g,15%). ¹H NMR (CD₃OD) δ: 1.58 (s, 3H), 1.66 (s, 3H), 2.22 (s, 3H), 2.38(m, 2H), 2.45 (m, 1H), 2.60 (m, 1H), 2.78 (dd, J=13.26, 8.21 Hz, 1H),2.99 (dd, J=13.52, 6.44 Hz, 1H), 3.15 (s, 1H), 3.35 (m, 1H), 3.75 (m,1H), 4.28 (d, J=11.12, 1H), 4.65 (m, 1H), 6.99 (t, J=7.01, 1H), 7.08 (t,J=7.58 Hz, 2H) 7.10-7.13 (m, 2H) 7.27 (d, J=5.31 Hz, 1H) 8.07 (d, J=5.31Hz, 1H). Anal. (C₂₆H₂₉N₈OSCl.0.5HOAc.0.5H₂O) C, H, N. HPLC: >95% purity.

Example 43-[(2,6-dichloropyrimidin-4-yl)amino]-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide

Preparation of Compound 4a:ethyl-3-amino-6,6-dimethyl-5-({[trans-2-phenylcyclopropyl]amino}carbonyl)-5,6-dihydropyrrolo[3,4-c]pyrazole-2(4H)-carboxylate

To a stirring mixture of 5-tert-butyl 2-ethyl3-amino-6,6-dimethylpyrrolo[3,4-c]pyrazole-2,5(4H,6H)-dicarboxylate(5.65 g, 17.4 mmol) in CH₂Cl₂ (20 ml) was added 4.0M HCl in dioxane (30ml). After the reaction mixture was concentrated in vacuo, a portion ofresidue (3.45 g, 11.6 mmol) was dissolved into a solution of DIPEA (8.1ml, 4 eq) in CH₂Cl₂ (50 ml). To the resulting solution was slowly addedto trans-2-phenylcyclopropyl isocyanate at 0° C. and the resultingmixture was stirred at 0° C. for 30 minutes then warmed up and stirredat room temperature for 1 h. The reaction mixture was diluted withCH₂Cl₂, and washed with saturated NaHCO₃, dried over sodium sulfate,concentrated in vacuo, purified by flash chromatography. Elution with60-80% EtOAc/hexane provided the compound 4a as a white solid (4.24 g,95%). ¹H NMR (CD₃OD) δ: 1.03-1.16 (m, 2H) 1.60 (d, J=3.79 Hz, 6H) 1.98(s, 1H) 2.71 (s, 1H) 4.12 (s, 1H) 7.01-7.10 (m, 3H) 7.13-7.21 (m, 2H)

Preparation of Compound 4b:3-amino-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide

To a stirring solution of ethyl3-amino-6,6-dimethyl-5-({[trans-2-phenylcyclopropyl]amino}carbonyl)-5,6-dihydropyrrolo[3,4-c]pyrazole-2(4H)-carboxylate(613 mg, 1.60 mmol) in MeOH (3 mL) was added 2N LiOH (1 ml, 1.25 eq).The resulting mixture was stirred under reflux for 4 hours, cooled, andconcentrated. The residue was partitioned between ethyl acetate andsaturated NaHCO₃, dried, and concentrated to give compound 4b as a whitesolid (0.42 g, 79%). ¹H NMR (CD₃OD) δ: 1.03-1.16 (m, 2H) 1.60 (d, J=3.79Hz, 6H) 1.98 (s, 1H) 2.71 (s, 1H) 4.12 (s, 2H) 7.01-7.10 (m, 3H)7.13-7.21 (m, 2H).

To a stirring solution of compound 4b (0.10 g, 0.32 mmol) in DMA (0.5mL) was added 2,4,6-trichloropyrimidine (0.041 ml, 1.1 eq) andtriethylamine (0.089 ml, 2 eq). The resulting mixture was heated to atemperature of 80° C. for 5 minutes in microwave reactor. The reactionmixture was purified by prep-HPLC to provide the compound 4 as a whitesolid (0.025 g, 17¹H NMR (CD₃OD) δ: 1.05-1.11 (m, 2H), 1.66 (d, J=3.54Hz, 6H), 1.95 (m, 1H), 2.68-2.72 (m, 1H), 4.42 (b, 2H), 7.05 (d, J=7.83Hz, 4H), 7.15 (t, J=7.58 Hz, 2H). Anal. (C₂₁H₂₁N₇OCl₂.0.4HOAc.0.2H₂O) C,H, N. HPLC: >95% purity.

Structure and Example # Chemical name, Analytical data and comments

5 N-[(1S)-2-(dimethylamino)-1-phenylethyl]-6,6-dimethyl-3-(thieno[3,2-d]pyrimidin-4-ylamino)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. ¹H NMR (400 MHz, CD₃OD) δ: 1.63 (s, 3 H),1.70 (s, 3 H), 2.38 (s, 6 H), 2.54 (dd, J = 12.63, 4.55 Hz, 1 H),2.80-2.95 (m, 1 H), 4.45-4.70 (m, 2 H), 4.94-5.08 (m, 1 H), 7.09- 7.41(m, 6 H), 7.95-8.09 (m, 1 H), 8.59 (s, 1 H). Anal. (C₂₄H₂₈N₈OS•0.75HOAc) C, H, N. Method of Example 1. [(2S)-2-isocyanato-2-phenyl ethyl]dimethylamine was used in place of trans-2- phenylcyclopropylisocyanate.

6 3-[(2-chlorothieno[3,2-d]pyrimidin-4-yl)amino]-N-[(1S)-2-(dimethylamino)-1-phenylethyl]-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. ¹H NMR (400 MHz, CD₃OD)δ: 1.62 (s, 3 H), 1.68 (s, 3 H), 2.38 (s, 6 H), 2.56-2.66 (m, 1 H),2.79-2.89 (m, 1 H), 4.69-4.73 (m, 1 H), 4.80-4.85 (m, 1 H), 4.93 (dd, J= 10.2, 4.4 Hz, 1 H), 7.07-7.38 (m, 6 H), 8.05 (d, J = 5.31 Hz, 1 H).Anal. (C₂₄H₂₇N₈OSCl•1.0 HOAc•0.6H₂O) C, H, N. Method of Example 2.[(2S)-2-isocyanato-2-phenylethyl] dimethylamine was used in place oftrans-2- phenylcyclopropyl isocyanate 2.

7 3-[(4-chloro-6-propoxy-1,3,5-triazin-2-yl)amino]-6,6-dimethyl-N-[trans-2-phenyl cyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (CD₃OD) δ: 0.94(t, J = 7.33 Hz, 3 H) 1.02- 1.13 (m, 2 H) 1.65 (d, J = 3.54 Hz, 6 H)1.68-1.77 (m, 2 H) 1.90-2.00 (m, 1 H) 2.65-2.73 (m, 1 H) 4.27 (t, J =6.19 Hz, 2 H) 4.39 (d, J = 23.75 Hz, 2 H) 6.99-7.08 (m, 3 H) 7.10-7.19(m, 2 H). Anal. (C₂₃H₂₇N₈O₂Cl•0.2HOAc•0.6H₂O) C, H, N. Method of Example4. 2,4-dichloro-6-propoxy-1,3,5 triazine was used in place of2,4,6-trichloropyrimidine while the reaction mixture was heated to 120°C.

8 3-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. ¹H NMR (CD₃OD) δ:1.02-1.16 (m, 2 H) 1.58-1.69 (m, 4 H) 1.88 (d, J = 4.04 Hz, 2 H)1.91-2.01 (m, 1 H) 2.63-2.75 (m, 1 H) 4.12 (d, 1 H) 4.36 (b, 1 H)6.98-7.09 (m, 3 H) 7.10-7.20 (m, 2 H). Anal.(C₂₀H₂₂N₉OCl•0.3HOAc•1.1H₂O)C, H, N. Method of example 4.2,4-dichloro-6-amino-1,3,5- triazine was used in place 2,4,6-trichloropyrimidine and the reaction mixture was heated to 120° C.

9 3-[(4,6-dichloro-1,3,5-triazin-2-yl)amino]-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. ¹H NMR (CD₃OD) δ: 1.02-1.14 (m, 2 H) 1.66(d, J = 3.54 Hz, 6 H) 1.91-1.99 (m, 1 H) 2.65-2.74 (m, 1 H) 4.44 (s, 2H) 7.00-7.08 (m, 3 H) 7.10-7.18 (m, 2 H). Anal.(C₂₀H₂₀N₈OCl₂•0.1HOAc•0.4H₂O) C, H, N. Method of example 4. 2,4,6-trichloro-1,3,5-triazine was used in place of 2,4,6-trichloropyrimidineand the reaction mixture was stirred at room temperature overnight.

10 3-[(4-chloro-6-methoxy-1,3,5-triazin-2-yl)amino]-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. ¹H NMR (CD₃OD) δ:1.02-1.16 (m, 2 H) 1.66 (d, J = 3.54 Hz, 6 H) 1.92-1.99 (m, 1 H) 2.70(b, 1 H) 3.92 (s, 3 H) 4.38 (d, J = 33.85 Hz, 2 H) 6.99-7.09 (m, 3 H)7.11-7.19 (m, 2 H). Anal. (C₂₁H₂₃N₈O₂Cl•0.1 HOAc•0.3H₂O) C, H, N. Methodof example 4. 2,4-dichloro-6-methoxy-1,3,5- triazine was used in placeof 2,4,6-trichloropyrimidine and the reaction mixture was stirred atroom temperature for 1 hr.

11 6,6-dimethyl-3-{[2-(methylthio)-6-(trifluoromethyl)pyrimidin-4-yl]amino}-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole- 5(1H)-carboxamide.¹H NMR (400 MHz, DMSO-d₆) δ ppm 1.00-1.09 (m, 1 H) 1.14-1.21 (m, 1 H)1.63 (d, J = 1.52 Hz, 6 H) 1.87-1.94 (m, 1 H) 2.69-2.79 (m, 1 H) 3.32(s, 3 H) 4.34 (s, 2 H) 6.12 (s, 1 H) 6.81 (s, 1 H) 7.07-7.17 (m, 3 H)7.24 (t, J = 7.58 Hz, 2 H) 10.52 (s, 1 H). Anal. (C₂₃H₂₄N₇OSF₃•0.6H₂O)C, H, N. Method of Example 4. 4-chloro-2-(methylthio)-6-(trifluoromethyl) pyrimidine was used in place of 2,4,6-trichloropyrimidine while the reaction mixture was stirred at 100° C.for 10 minutes in microwave reactor.

12 3-[(2-chloroquinazolin-4-yl)amino]-6,6-dimethyl-N-[(trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. ¹H NMR (CD₃OD) δ: 1.04-1.15 (m, 2 H) 1.70(d, J = 3.28 Hz, 6 H) 1.92-2.00 (m, 1 H) 2.67-2.77 (m, 1 H) 4.56 (s, 2H) 6.08 (s, 1 H) 7.00-7.09 (m, 3 H) 7.10-7.19 (m, 2 H) 7.54 (t, J = 7.45Hz, 1 H) 7.64 (d, J = 8.34 Hz, 1 H) 7.79 (t, J = 7.33 Hz, 1 H) 8.26 (d,J = 8.08 Hz, 1 H). Anal. (C₂₅H₂₄N₇OCl•0.2HOAc•0.5H₂O) C, H, N. Method ofExample 4. 2,4-dichloroquinazoline was used in place of2,4,6-trichloropyrimidine while the reaction mixture was stirred at 160°C. for 10 minutes in a microwave reactor.

Example 133-({4-[(2S)-2-(aminocarbonyl)pyrrolidin-1-yl]-6-chloro-1,3,5-triazin-2-yl}amino)-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide

To a stirring mixture of Compound 9 (0.039 g, 0.085 mmol) and DIEPA(0.030 ml, 2 eq) in THF (1 ml) was added L-prolinamide (9.7 mg, 1 eq).The resulting mixture was stirred at room temperature for 1 hr andpurified by prep-HPLC to provide the compound 13 as a white solid (23mg). ¹H NMR (CD₃OD) δ: 1.01-1.16 (m, 2H) 1.57-1.71 (m, 6H) 1.88-2.09 (m,4H) 2.12-2.35 (m, 1H) 2.65-2.76 (m, 1H) 3.50-3.78 (m, 2H) 4.27-4.53 (m,3H) 6.99-7.08 (m, 3H) 7.10-7.19 (m, 2H). Anal.(C₂₆H₂₉N₁₀O₂Cl.0.4HOAc.1.3H₂O) C, H, N. HPLC: >95% purity.

Example 143-[(4,6-dimethoxy-1,3,5-triazin-2-yl)amino]-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide

To a stirring solution of compound 4b (0.150 g, 0.48 mmol) in DMA (1 mL)was added 2,4,6-trichlorotriazene (98 mg, 1.1 eq) and DIPEA (0.168 ml, 2eq). The resulting mixture was stirred at room temperature for 1 hr. Tothe reaction mixture was added 25% NaOMe in methanol (0.549 ml, 5 eq).The resulting mixture was stirred at room temperature for 2 hours andpurified by prep-HPLC to provide compound 14 as a white solid (45 mg).¹H NMR (CD₃OD) δ: 1.02-1.14 (m, 2H) 1.65 (d, J=3.79 Hz, 6H) 1.91-1.99(m, 1H) 2.64-2.72 (m, 1H) 3.89 (s, 6H) 4.32 (s, 2H) 6.99-7.08 (m, 3H)7.09-7.18 (m, 2H). Anal. (C₂₂H₂₆N₈O₃.0.2HOAc.0.6H₂O) C, H, N. HPLC: >95%purity.

Structure and Example # Chemical name, Analytical data and Comments

15 3-{[4-chloro-6-(dimethylamino)-1,3,5-triazin-2-yl]amino}-6,6-dimethyl-N-[trans-2-phenyl cyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. ¹H NMR (CD₃OD) δ: 1.02-1.15 (m, 2 H)1.64 (d, J = 3.79 Hz, 6 H) 1.91-1.98 (m, 1 H) 2.65-2.74 (m, 1 H) 3.07(s, 6 H) 4.33 (s, 2 H) 7.00-7.09 (m, 3 H) 7.10-7.18 (m, 2 H). Anal.(C₂₂H₂₆N₉OCl•0.2HOAc•0.1H₂O) C, H, N. Method of Example 14, DMA was usedin place of NaOMe.

Example 163-({4-[(2S)-2-(hydroxymethyl)pyrrolidin-1-yl]-6-methoxy-1,3,5-triazin-2-yl}amino)-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide

To a stirring solution of compound 4b (0.152 g, 0.488 mmol) in DMA (2mL) was added 2,4-dichloro-6-methoxy-1,3,5-triazine (92 mg, 1.05 eq) andDIPEA (0.170 ml, 2 eq). The resulting mixture was stirred at roomtemperature for 1 hr. To the reaction mixture was addedpyrrolidin-2-ylmethanol (0.072 ml, 1.5 eq). The resulting mixture wasstirred at room temperature for 1 hr and purified by prep-HPLC toprovide compound 16 as a white solid (64.9 mg). ¹H NMR (CD₃OD) δ:1.02-1.14 (m, 2H) 1.63 (d, J=3.54 Hz, 6H) 1.84 (b, 1H) 1.90-2.00 (m, 4H)2.66-2.74 (m, 1H) 3.48-3.71 (m, 4H) 3.84 (s, 3H) 4.10-4.22 (m, 1H) 4.28(s, 2H) 7.00-7.08 (m, 3H) 7.11-7.18 (m, 2H). Anal.(C₂₆H₃₃N₉O₃.0.1HOAc.0.7H₂O) C, H, N. HPLC: >95% purity.

Struture and Example # Chemical name, Analytical data and comments

17 3-[(4-methoxy-6-{(2S)-2-[(methylamino)carbonyl]pyrrolidin-1-yl}-1,3,5-triazin-2-yl)amino]-6,6-dimethyl-N-[(trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. ¹H NMR (CD₃OD) δ: 1.03-1.16 (m, 2 H)1.64 (d, J = 3.79 Hz, 6 H) 1.89-2.04 (m, 4 H) 2.14-2.28 (m, 1 H) 2.58-2.67 (m, 3 H) 2.71 (s, 1 H) 3.54-3.74 (m, 2 H) 3.82 (d, J = 37.14 Hz, 3H) 4.26 (d, J = 21.73 Hz, 2 H) 4.36-4.48 (m, 1 H) 7.01-7.09 (m, 3 H)7.11-7.19 (m, 2 H). Anal. (C₂₇H₃₄N₁₀O₃•0.2HOAc•0.9H₂O) C, H, N. Methodof Example 16 using N-methyl-L-prolinamide in place ofpyrrolidin-2-ylmethanol.

18 3-{[4-methoxy-6-(2-methylaziridin-1-yl)-1,3,5-triazin-2-yl]amino}-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. ¹H NMR (CD₃OD) δ:1.04-1.14 (m, 5 H) 1.64 (d, J = 3.79 Hz, 6 H) 1.93-2.01 (m, 1 H)2.66-2.75 (m, 1 H) 3.26- 3.46 (m, 2 H) 3.79-3.89 (m, 4 H) 4.30 (s, 2 H)6.99-7.09 (m, 3 H) 7.15 (t, J = 7.45 Hz, 2 H). Anal.(C₂₄H₂₉N₉O₂•0.2HOAC•1.6H₂O) C, H, N. Method of Example 16 using2-methylaziridine instead of pyrrolidin-2-ylmethanol

19 3-[(4-methoxy-6-pyrrolidin-1-yl-1,3,5-triazin-2-yl)amino]-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. ¹H NMR (CD₃OD) δ:1.03-1.14 (m, 2 H) 1.64 (d, J = 3.79 Hz, 6 H) 1.88-2.00 (m, 5 H)2.65-2.74 (m, 1 H) 3.43- 3.56 (m, 4 H) 3.84 (s, 3 H) 4.28 (s, 2 H)6.99-7.10 (m, 3 H) 7.10-7.19 (m, 2 H). Anal. (C₂₅H₃₁N₉O₂•0.2HOAc•0.6H₂O)C, H, N. Method of Example 16. Pyrrolidine was used in place ofpyrrolidin-2-ylmethanol.

20 3-({4-[(2S)-2-cyanopyrrolidin-1-yl]-6-methoxy-1,3,5-triazin-2-yl}amino)-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole- 5(1H)-carboxamide.¹H NMR (CD₃OD) δ: 1.02-1.15 (m, 2 H) 1.64 (d, J = 3.54 Hz, 6 H)1.91-2.01 (m, 1 H) 2.01-2.14 (m, 2 H) 2.21- 2.80 (m, 2 H) 2.65-2.73 (m,1 H) 3.46-3.57 (m, 1 H) 3.62-3.72 (m, 1 H) 3.89 (d, J = 13.39 Hz, 3 H)4.30 (s, 2 H) 4.87 (t, J = 5.18 Hz, 1 H) 7.00-7.08 (m, 3 H) 7.11-7.19(m, 2 H). Anal. (C₂₆H₃₀N₁₀O₂•0.3HOAc•0.1H₂O) C, H, N. Method of Example16. (2S)-pyrrolidine-2-carbonitrile was used in place ofpyrrolidin-2-ylmethanol.

21 3-({4-[(2S)-2-(amino carbonyl)pyrrolidin-1-yl]-6-methoxy-1,3,5-triazin-2-yl}amino)-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)- carboxamide. ¹HNMR (CD₃OD) δ: 1.02-1.16 (m, 2 H) 1.63 (s, 6 H) 1.90-2.08 (m, 4 H) 2.25(s, 1 H) 2.71 (s, 1 H) 3.56-3.75 (m, 2 H) 3.84 (d, J = 21.73 Hz, 3 H)4.21-4.37 (m, 2 H) 4.44 (d, J = 9.09 Hz, 1 H) 7.00-7.08 (m, 3 H)7.10-7.19 (m, 2 H). Anal. (C₂₆H₃₂N₁₀O₃•0.2HOAc•0.9H₂O) C, H, N. Methodof Example 16 using L-prolinamide (2eq) instead ofpyrrolidin-2-ylmethanol.

Example 223-{[4-[(2S)-2-(aminocarbonyl)pyrrolidin-1-yl]-6-(dimethylamino)-1,3,5-triazin-2-yl]amino}-6,6-dimethyl-N-[(trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide

To a stirring solution of compound 4b (0.103 g, 0.33 mmol) in DMA (1 mL)was added 2,4,6-trichloropyrimidine (55 mg, 0.9 eq) and DIPEA (0.115 ml,2 eq). The resulting mixture was stirred at room temperature for 1 hr.To the reaction mixture was added L-prolinamide (34 mg, 0.9 eq). Afterthe resulting mixture was stirred at room temperature for 2 hours, 2Ndimethylamine (0.330 ml, 2 eq) was added into reaction mixture. Theresulting mixture was stirred at 60° C. for 1 hour and purified byprep-HPLC to provide compound 22 as a white solid (74.8 mg). ¹H NMR(CD₃OD) δ: 1.04-1.17 (m, 2H) 1.62 (d, J=3.28 Hz, 6H) 1.92 (m, 4H) 2.22(s, 1H) 2.67-2.76 (m, 1H) 3.03 (d, 6H) 3.55-3.75 (m, 2H) 4.21 (d,J=14.15 Hz, 2H) 4.35-4.43 (m, 1H) 7.01-7.09 (m, 3H) 7.11-7.19 (m, 2H).Anal. (C₂₇H₃₅N₁₁O₂.0.3HOAc.1.0H₂O) C, H, N. HPLC: >95% purity.

Structure and Example # Chemical name, Analytical data and comments

3-({4-(dimethylamino)-6-[(2S)-2-(hydroxymethyl)pyrrolidin-1-yl]-1,3,5-triazin-2-yl}amino)-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole- 5(1H)-carboxamide.¹H NMR (CD₃OD) δ: 1.02-1.16 (m, 2 H) 1.62 (d, J = 3.79 Hz, 6 H)1.91-2.02 (m, 4 H) 2.63-2.74 (m, 1 H) 3.04 (s, 6 H) 3.43-3.68 (m, 4 H)4.12-4.31 (m, 4 H) 7.01-7.09 (m, 3 H) 7.11-7.20 (m, 2 H). Anal.(C₂₇H₃₆N₁₀O₂• 0.2HOAc•0.8H₂O) C, H, N. Method of Example 22 using(2S)-pyrrolidin-2-ylmethanol in place of L-prolinamide

Example 243-[(4,6-dimethylpyrimidin-2-yl)amino]-6,6-dimethyl-N-[(trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide

A re-sealable tube was charged with compound 4b (0.115 g, 0.30 mmol),2-chloro-4,6-dimethyl-pyrimidine (0.043 g, 1 eq), Pd(OAc)₂ (1.3 mg, 0.02eq), DPPP (5.0 mg, 0.04 eq), CsCO₃ (137 mg, 1.4 eq) and DME (1 ml). Thetube was capped and carefully subjected to three cycles ofevacuation-backfilling with N₂. The resulting mixture was stirred at150° C. for 10 minutes in microwave reactor, filtered, and purified byprep-HPLC to provide compound 24 as a white solid (12 mg). ¹H NMR(CD₃OD) δ: 1.03-1.14 (m, 2H) 1.65 (d, J=3.79 Hz, 6H) 1.92-2.00 (m, 1H)2.30 (s, 6H) 2.68-2.75 (m, 1H) 4.33 (s, 2H) 6.58 (s, 1H) 7.00-7.09 (m,3H) 7.11-7.19 (m, 2H). Anal. (C₂₃H₂₇N₇O.0.3HOAc.0.3H₂O) C, H, N.HPLC: >95% purity.

Structure and Example # Chemical name, Analytical data and comments

3-[(4-trifluoromethylpyridin-2-yl)amino]-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. ¹H NMR (CD₃OD) δ: 1.01-1.17 (m, 2 H) 1.65(d, J = 3.79 Hz, 6 H) 1.93-2.01 (m, 1 H) 2.64-2.74 (m, 1 H) 4.35 (s, 2H) 6.90 (d, J = 5.31 Hz, 1 H) 6.98 (s, 1 H) 7.00-7.09 (m, 3 H) 7.10-7.18(m, 2 H) 8.32 (d, J = 5.31 Hz, 1 H). Anal. (C₂₃H₂₃N₆OF₃•0.1HOAc•0.4H₂O)C, H, N. Method of Example 24. 4,5-bis (diphenylphosphino)-9,9-dimethylxanthene was used in place of DPPP and 2-chloro-4-trifluoromethyl-pyridine was used in place of 2-chloro-4,6-dimethyl-pyrimidine.

3-[(4-trifluoromethyl-6-methylpyridin-2-yl)amino]-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. ¹H NMR (CD₃OD) δ:1.02-1.14 (m, 2 H) 1.65 (d, J = 3.54 Hz, 6 H) 1.90-2.00 (m, 1 H) 2.45(s, 3 H) 2.66-2.73 (m, 1 H) 4.34 (s, 2 H) 6.79 (s, 2 H) 6.99-7.09 (m, 3H) 7.10- 7.18 (m, 2 H). Anal. (C₂₄H₂₅N₆OF₃•0.2HOAc•0.1H₂O) C, H, N.Method of Example 24. 4,5-bis (diphenylphosphino)-9,9- dimethylxanthenewas used in place of DPPP and 2-chloro-4-trifluoromethyl-6-methylpyridine was used in place of2-chloro-4,6-dimethyl-pyrimidine.

3-[(6-trifluoromethylpyridin-2-yl)amino]-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. ¹H NMR (CD₃OD) δ: 1.05-1.13 (m, 2 H) 1.65(d, J = 3.79 Hz, 6 H) 1.89-1.98 (m, 1 H) 2.67-2.76 (m, 1 H) 4.40 (s, 2H) 6.96 (d, J = 8.08 Hz, 1 H) 7.01-7.11 (m, 4 H) 7.11- 7.19 (m, 2 H)7.66 (t, J = 7.45 Hz, 1 H). Anal. (C₂₃H₂₃N₆OF₃•0.1HOAc•0.3H₂O) C, H, N.Method of Example 24. 4,5-bis (diphenylphosphino)-9,9- dimethylxanthenewas used in place of DPPP and 2- chloro-6-trifluoromethylpyridine wasused in place of 2- chloro-4,6-dimethyl-pyrimidine.

6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-3-{[4-(trifluoromethyl)pyrimidin-2-yl]amino}-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. ¹H NMR (CD₃OD) δ:1.03-1.15 (m, 2 H) 1.65 (d, J = 3.54 Hz, 6 H) 1.93-2.00 (m, 1 H)2.67-2.76 (m, 1 H) 4.40 (b, 2 H) 6.98-7.11 (m, 4 H) 7.12-7.19 (m, 2 H)8.67 (b, 1 H). Anal. (C₂₂H₂₂N₇OF₃•0.1 H₂O) C, H, N. Method of Example24. 2-chloro-4-trifluoromethyl- pyrimidine was used in place of2-chloro-4,6-dimethyl- pyrimidine.

3-[(4-cyanopyridin-2-yl)amino]-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole- 5(1H)-carboxamide.¹H NMR (CD₃OD) δ: 1.02-1.15 (m, 2 H) 1.65 (d, J = 3.79 Hz, 6 H)1.94-2.01 (m, 1 H) 2.66-2.74 (m, 1 H) 4.34 (s, 2 H) 6.91 (d, J = 4.80Hz, 1 H) 6.99-7.08 (m, 4 H) 7.11- 7.19 (m, 2 H) 8.29 (d, J = 5.05 Hz, 1H). Anal. (C₂₃H₂₃N₇O•0.3HOAc•0.4H₂O) C, H, N. Method of Example 24.4,5-bis (diphenylphosphino)-9,9- dimethylxanthene was used in place ofDPPP and 2- chloro-4-cyanopyridine was used in place of 2-chloro-4,6-dimethyl-pyrimidine.

Example 306,6-dimethyl-N-[trans-2-phenylcyclopropyl]-3-{[2-(trifluoromethyl)pyrimidin-4-yl]amino}-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide

Preparation of Compound 30a: 4-chloro-2-(trifluoromethyl)pyrimidine

To 2-(trifluoromethyl)pyrimidin-4-ol (2 g, 12 mmol) was added POCl₃ (15mL). It was refluxed overnight. The solvent was removed. 1N NaOH wasadded to the reaction mixture slowly until pH=10. The mixture wasextracted with DCM (3×80 mL). Combined DCM layer dried over Na₂SO₄ andtaken to dryness. The crude product was vacuum distilled.4-chloro-2(trifluoromethyl)pyrimidine was obtained as a clear oil.(94%). ¹H NMR (400 MHz, DCM) δ: 7.60 (d, J=3 Hz, 1H), 8.80 (d, J=3 Hz,1H),

To a mixture of compound 30a (137 mg, 0.75 mmol) and compound 4b (235mg, 0.75 mmol) in IPA (1 mL), was added TEA (210 mL, 1.5 mmol). Thereaction was heated in microwave oven at 140° C. for 20 min. HPLCyielded the desired product as a white powder (28 mg, 6.4%). ¹H NMR (400MHz, DMSO) δ: 1.04 (m, 2H), 1.62 (m, 6H), 1.88 (m, 1H), 2.73 (m, 1H),4.34 (m, 2H), 7.09-7.37 (m, 6H), 8.41 (s, 1H). Anal.(C₂₂H₂₂N₇OF₃.0.52TFA.0.54H₂O) C, H, N. APCI-MS: [M+H] 458.

Example 316,6-dimethyl-3-[(2-methylthieno[2,3-d]pyrimidin-4-yl)amino]-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide

A mixture of compound 4b (83 mg, 0.267 mmol) and compound 31c(4-chloro-2-methylthieno[2,3-d]pyrimidine, prepared following samemethod compound 32c was prepared, 115 mg, 2 eq), was added an aqueoussolution of acetic acid (50% v/v, 1 ml). The resulted mixture was heatedand stirred at 100° C. for 1 hour. Preparative HPLC purification givethe title compound as a white solid (34 mg, 27% yield). ¹H NMR (CD₃OD)δ: 1.03-1.13 (m, 2H), 1.69 (s, 3H), 1.70 (s, 3H), 1.90-1.99 (m, 1H),2.66 (s, 3H), 2.67-2.75 (m, 1H), 4.46 (s, 2H), 6.98-7.19 (m, 5H),7.55-7.61 (m, 1H) 7.63-7.69 (m, 1H), 8.20 (d, J=6.06 Hz, 1H). LCMS(APCI, M+H⁺): 460.1. Anal. (C₂₄H₂₆N₇OS.1.51TFA.0.15H₂O): C, H, N.HPLC-UV Detection: 95% purity.

Example 326,6-Dimethyl-3-[(2-methylthieno[3,2-d]pyrimidin-4-yl)amino]-N-[(1R,2S)-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide

Preparation of Compound 32b: 2-Methylthieno[3,2-d]pyrimidin-4(3H)-one

Methyl 3-(acetylamino)thiophene-2-carboxylate (32a, 3.00 g, 15.08 mmol)was suspended in 30% NH₄OH (43 mL) in a sealed tube. The reaction wasstirred at 120° C. for 5 hours and then overnight at ambienttemperature. The reaction was brought to pH 8-9 with concentrated HCl.The resulting white precipitate was filtered and washed with water, thendried to give compound 32b (1.56 g, 62%) as a white solid. Compound 32bwas used without further purification. ¹H NMR (400 MHz, DMSO-d₆) δ 2.35(s, 3H), 7.30 (d, J=5.3 Hz, 1H), 8.12 (d, J=5.3 Hz, 1H), 12.38 (s, 1H).LCMS 167 (M+H).

Preparation of Compound 32c: 4-Chloro-2-methylthieno[3,2-d]pyrimidine

2-Methylthieno[3,2-d]pyrimidin-4(3H)-one (compound 32b, 0.18 g, 1.13mmol) was heated to 110° C. in POCl₃ overnight. The solvent was removedunder reduced pressure, and the residue was neutralized with saturatedNaHCO₃ solution. The product was extracted into CH₂Cl₂, and the organicphase separated, washed with brine, and dried (MgSO₄). After removal ofthe solvent compound 32c was obtained as a yellow-orange solid (0.21 g,88%). Compound 32c was used without further purification. R_(f)=0.16(10% EtOAc/hexane). ¹H NMR (400 MHz, DMSO-d₆): δ 2.71 (s, 3H), 7.66 (d,J=5.6 Hz, 1H), 8.54 (d, J=5.6 Hz, 1H). LCMS 185 (M+H). Anal.(C₂₃H₂₉N₈OF.0.35H₂O.0.35 hexane) C, H, N. HPLC>98% purity.

Preparation of Enantiomer 32d(3-amino-6,6-dimethyl-N-((1R,2S)-2-phenylcyclopropyl)pyrrolo[3,4-c]pyrazole-5(1H,4H,6H)-carboxamide)and enantiomer 32e (3-amino-6,6-dimethyl-N-((1S,2R)-2-phenylcyclopropyl)pyrrolo[3,4-c]pyrazole-5(1H,4H,6H)-carboxamide)

An enantioseparation purification method was developed for compound 4busing supercritical fluid chromatography (SFC) technology, withsupercritical carbon dioxide providing the bulk of the mobile phase. Theseparation and isolation of enantiomers was carried out on a Berger SFCMultiGram™ Purification System (Mettler Toledo AutoChem, Inc.). Thepreparative chromatography conditions used to separate the enantiomersconsisted of a (S,S) Whelk-O 1 (Regis Technologies, Inc.), 10/100 FEC,250×21.1 mm column as the chiral stationary phase. Column temperaturewas maintained at 35° C. The mobile phase used was supercritical CO₂with 35% methanol as the modifier, maintained isocratically at a flowrate of 55 mL/min and a constant pressure of 140 bar. Sample wassolubilized in methanol, and a column loadability of 50 mg per 1 mLinjection was attained, and the total run time for each injection was7.0 minutes. Retention times for the two enantiomers were 4.3 and 5.8minutes, respectively. The specific optical rotation, [α]_(D), for thepure enantiomers was determined to be −126.7° for Enantiomer 32d and+124.4° for Enantiomer 32e

Preparation of Title Compound 32

4-Chloro-2-methylthieno[3,2-d]pyrimidine (0.091 g, 0.49 mmol) and chiralaminopyrazole enantiomer 32d (0.10 g, 0.33 mmol) were mixed in 1:1HOAc/H₂O (1.40 mL) and heated to 100° C. for 1 hour. The material wasthen purified directly by preparative HPLC to give title compound 32 asa white solid (0.136 g, 65%). Mp>148° C. (dec). ¹H NMR (400 MHz, CH₃OD):δ 1.15-1.19 (m, 2H), 1.79 (s, 3H), 1.80 (s, 3H), 2.01-2.05 (m, 1H),2.76-2.78 (m, 1H), 2.80 (s, 3H), 4.50-4.51 (m, 2H), 7.11-7.15 (m, 3H),7.22-7.25 (m, 2H), 7.46 (d, J=5.5 Hz, 1H), 8.44 (d, J=5.3 Hz, 1H). LCMS460 (M+H). Anal. (C₂₄H₂₆N₇OS.1.50 TFA.0.25H₂O) C,H,N. HPLC>99% purity.

Structure and Example # Chemical name, Analytical data and comments

3-[(2-cyanopyrimidin-4-yl)amino]-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (CD₃OD) δ: 1.03-1.15 (m, 2 H), 1.66(s, 6 H), 1.75-2.01 (m, 1 H), 2.51-2.75 (m, 1 H), 4.59 (s, 2 H),6.81-6.96 (b, 1 H), 7.00-7.07 (m, 3 H), 7.10-7.17 (m, 2 H), 8.20 (d, J =6.06 Hz, 1 H). LCMS (APCI, M + H⁺): 415.1. HPLC-UV Detection: >95%purity Method of Example 31 using 4-chloropyrimidine-2- carbonitrile inplace of 31c.

6,6-dimethyl-3-[(2-methyl-6-morpholin-4-ylpyrimidin-4-yl)amino]-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (400 MHz, DMSO)δ: 1.04-1.21 (m, 2 H), 1.63 (s, 6 H), 2.42 (s, 3 H), 2.75 (m, 1 H), 4.28(m, 2 H), 6.05 (s, 1 H), 6.31 (m, 1 H) 7.09-7.27(m, 5H). Anal.(C₂₆H₃₂N₈O₈•1.6TFA•2.2H₂O) C, H, N. APCI- MS: [M + H] 489. Method ofExample 31 using 4-chloro-2-methyl-6- morpholinopyrimidine in place of31c.

6,6-dimethyl-3-[(6-methylthieno[2,3-d]pyrimidin-4-yl)amino]-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (CD₃OD) δ:1.04-1.17 (m, 2 H), 1.69 (s, 3 H), 1.70 (s, 3 H), 1.95-2.01 (m, 1 H),2.57 (s, 3 H), 2.67- 2.74 (m, 1 H) 4.45 (s, 2 H), 7.01-7.08 (m, 3 H),7.11- 7.18 (m, 2 H), 7.32 (s, 1 H), 8.52 (s, 1 H). LCMS (APCI, M + H⁺):460.3. Anal. (C₂₄H₂₅N₇OS•1.34TFA•0.47H₂O): C, H, N. HPLC-UV Detection:94% purity. Method of Example 31 using 4-chloro-6-methylthieno[2,3-d]pyrimidine in place of 31c.

6,6-dimethyl-3-[(9-methyl-9H-purin-6-yl)amino]-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (CD₃OD) δ: 1.01-1.17 (m, 2 H), 1.69(s, 3 H), 1.70 (s, 3 H), 1.94-2.03 (m, 1 H), 2.65-2.74 (m, 1 H), 3.83(s, 3 H), 4.43 (s, 2 H), 6.98-7.09 (m, 3 H), 7.11- 7.18 (m, 2 H), 8.21(s, 1 H), 8.53 (s, 1 H). LCMS (APCI, M + H⁺): 444.2. Anal.(C₂₃H₂₅N₉O•1.15TFA•0.59H₂O): C, H, N. HPLC-UV Detection: 90% purity.Method of Example 31 using 6-chloro-9-methyl-9H- purine in place of 31c.

3-[(2-ethoxy-5-fluoropyrimidin-4-yl)amino]-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (CD₃OD) δ:1.02-1.12 (m, 2 H), 1.30 (t, J = 10.21 Hz, 3H), 1.66 (s, 3 H), 1.67 (s,3 H), 1.90- 1.99 (m, 1 H), 2.65-2.72 (m, 1 H), 4.29 (q, J = 7.07 Hz, 2H), 4.35 (s, 2 H), 6.99-7.07 (m, 3 H), 7.11-7.19 (m, 2 H), 8.08 (d, J =3.39 Hz, 1 H). LCMS (APCI, M + H⁺): 452.2. Anal.(C₂₃H₂₆FN₇O₂•1.22TFA•0.59H₂O): C, H, N. HPLC-UV Detection: 89% purity.Method of Example 31 using 4-chloro-2-ethoxy-5- fluoropyrimidine inplace of 31c.

3-{[6-(dimethylamino)-2-methylpyrimidin-4-yl]amino}-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (400 MHz, DMSO)δ: 1.06-1.21 (m, 2 H), 1.62 (s, 6 H), 2.35 (s, 3 H), 2.67 (m, 1 H), 3.28(m, 7 H), 4.41 (s, 2 H), 7.10-7.27(m, 5H). APCI-MS: [M + H] 447. Methodof Example 31 using 6-chloro-N,N,2- trimethylpyrimidin-4-amine in placeof 31c.

6,6-dimethyl-3-[(2-methylpyrimidin-4-yl)amino]-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (400 MHz, DMSO) δ: 1.03 (m, 2 H),1.87 (s, 3 H), 2.09 (s, 3H), 2.40 (s, 3 H), 4.00 (m, 2 H), 5.30 (s, 2H), 6.49 (m, 1H) 7.09-7.27(m, 7H), 8.18 (m, 1 H),. APCI-MS: [M + H] 404.Method of Example 31 using 4-chloro-2- methylpyrimidine in place of 31c.

3-[(2-cyanoquinazolin-4-yl)amino]-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide R_(f) = 0.11 (7% methanolic NH₃/CHCl₃). ¹HNMR (400 MHz, CD₃OD): δ 1.15-1.25 (m, 2H), 1.79 (s, 6H), 2.06-2.11 (m,1H), 2.80-2.84 (m, 1H), 4.76-4.77 (m, 2H), 7.11-7.15 (m, 3H), 7.22-7.25(m, 2H), 7.75-7.79 (m, 1H), 7.88-7.90 (m, 1H), 7.95-7.98 (m, 1H), 8.41(d, J = 7.5 Hz, 1H). LCMS 465 (M + H). Anal. (C₂₆H₂₄N₈O•0.70 H₂O•0.05hexane) C, H, N. Method of Example 31 using 4-Chloroquinazoline-2-carbonitrile 40c in place of 31c.

6,6-dimethyl-3-[(2-methylthieno[3,2-d]pyrimidin-4-yl)amino]-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (400 MHz, CD₃OD):δ 1.15-1.19 (m, 2H), 1.79 (s, 3H), 1.80 (s, 3H), 2.01-2.05 (m, 1H),2.76-2.78 (m, 1H), 2.80 (s, 3H), 4.50-4.51 (m, 2H), 7.11-7.15 (m, 3H),7.22-7.25 (m, 2H), 7.46 (d, J = 5.5 Hz, 1H), 8.44 (d, J = 5.3 Hz, 1H).LCMS 460 (M + H). Anal. (C₂₄H₂₅N₇OS•1.70 TFA•0.25 H₂O) C, H, N. Methodof Example 31 using 4-Chloro-2- methylthieno[3,2-d]pyrimidine (32c) inplace of 31c.

6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-3-(thieno[2,3-d]pyrimidin-4-ylamino)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (CD₃OD) δ:1.03-1.17 (m, 2 H), 1.69 (s, 3 H), 1.70 (s, 3 H), 1.93-2.03 (m, 1 H),2.67-2.75 (m, 1 H), 4.47 (s, 2 H), 7.01-7.11 (m, 3 H), 7.15 (t, J = 7.58Hz, 2 H), 7.44 (d, J = 5.56 Hz, 1 H), 8.30 (d, J = 5.30 Hz, 1 H), 8.77(s, 1 H). LCMS (APCI, M + H⁺): 446.2. Anal.(C₂₃H₂₃N₇OS•1.86TFA•0.29H₂O•0.49TEA): C, H, N. HPLC-UV Detection: 87%purity Method of Example 31 using 4-chlorothieno[2,3- d]pyrimidine inplace of 31c.

3-[(5-fluoro-2-methoxypyrimidin-4-yl)amino]-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. ¹H NMR (400 MHz,CD₃OD): δ 1.13-1.21 (m, 2H), 1.75 (s, 3H), 1.76 (s, 3H), 2.01-2.06 (m,1H), 2.76-2.79 (m, 1H), 3.96 (s, 3H), 4.42-4.43 (m, 2H), 7.11-7.14 (m,3H), 7.21-7.25 (m, 2H), 8.13 (d, J = 3.8 Hz, 1H). LCMS 438 (M + H).Anal. (C₂₂H₂₄N₇O₂F•1.00 TFA•0.10 H₂O) C, H, N. Method of Example 31using 4-Chloro-5-fluoro-2- methoxypyrimidine 43b in place of 31c.

3-[(5-fluoro-2-methylpyrimidin-4-yl)amino]-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. ¹H NMR (400 MHz,CD₃OD): δ 1.16-1.21 (m, 2H), 1.76 (s, 3H), 1.77 (s, 3H), 2.01-2.06 (m,1H), 2.67 (s, 3H), 2.77-2.80 (m, 1H), 4.52-4.53 (m, 2H), 7.11-7.15 (m,3H), 7.22-7.26 (m, 2H), 8.43 (d, J = 5.0 Hz, 1H). LCMS 422 (M + H).Anal. (C₂₂H₂₄N₇OF•1.50 TFA) C, H, N. HPLC = 87% purity. Method ofExample 31 using 4-Chloro-5-fluoro-2- methylpyrimidine 44b in place of31c.

6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-3-{[2-(trifluoromethyl)thieno[3,2-d]pyrimidin-4-yl]amino}-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (400 MHz, MeOD) δppm: 1.18 (t, 2 H), 1.79 (d, J = 3.27 Hz, 6 H), 1.98-2.06 (m, 1 H),2.77-2.82 (m, 1 H), 4.52 (s, 2 H), 7.11-7.16 (m, 3 H), 7.24 (t, J = 7.68Hz, 2 H), 7.54 (d, J = 5.04 Hz, 1 H), 8.24 (d, J = 4.78 Hz, 1 H). Anal.C₂₄H₂₂N₇OF₃S•0.5HOAc•0.1H₂O) C, H, N. HPLC: >95% purity. Method ofExample 31 using 4-chloro-2- (trifluoromethyl)thieno[3,2-d]pyrimidine45c in place of 31c.

6,6-dimethyl-3-[(2-methylthieno[2,3-d]pyrimidin-4-yl)amino]-N-[(1R,2S)-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (CD₃OD) δ:1.02-1.15 (m, 2 H), 1.68 (s, 3 H), 1.69 (s, 3 H), 1.90-1.99 (m, 1 H),2.56 (s, 3 H), 2.66- 2.76 (m, 1 H), 4.43 (s, 2 H), 6.99-7.10 (m, 3 H),7.14 (t, J = 6.06 Hz, 2 H), 7.40 (d, J = 6.06 Hz, 1 H), 7.51 (d, J =6.06 Hz, 1 H). LCMS (APCI, M + H⁺): 460.2. Anal.(C₂₄H₂₅N₇OS•0.76H₂O•0.22HOAc): C, H, N. HPLC-UV Detection: 94% purity.Method of Example 32 using 4-chloro-2- methylthieno[2,3-d]pyrimidine(31c) in place of 32c.

6,6-dimethyl-3-[(2-methylthieno[2,3-d]pyrimidin-4-yl)amino]-N-[(1S,2R)-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (CD₃OD) δ:1.02-1.17 (m, 2 H), 1.68 (s, 3 H), 1.69 (s, 3 H), 1.92-2.02 (m, 1 H),2.56 (s, 3 H), 2.67- 2.79 (m, 1 H), 4.43 (s, 2 H), 7.01-7.11 (m, 3 H),7.14 (t, J = 8.08 Hz, 2 H), 7.40 (d, J = 6.06 Hz, 1 H), 7.51 (d, J =6.06 Hz, 1 H). LCMS (APCI, M + H⁺): 460.2. Anal. Calcd for(C₂₄H₂₅N₇OS•0.82H₂O•0.22HOAc): C, H, N. HPLC-UV Detection: 93% purityMethod of Example 32 using 4-chloro-2- methylthieno[2,3-d]pyrimidine(31c) in place of 32c, and 32e in place of 32d.

3-[(5-fluoropyrimidin-4-yl)amino]-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (400 MHz, CD₃OD): δ 1.13-1.25 (m,2H), 1.75 (s, 3H), 1.76 (s, 3H), 2.04-2.10 (m, 1 H), 2.76-2.80 (m, 1H),4.50-4.53 (m, 2H), 7.11-7.25 (m, 5H), 8.25 (s, 1H), 8.51 (s, 1 H). LCMS408 (M + H). HPLC >99% purity. Method of Example 31 using 4-Chloro-5-fluoropyrimidine 48b in place of 31c.

6,6-dimethyl-3-[(7-methylthieno[3,2-d]pyrimidin-4-yl)amino]-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (400 MHz, DMSO)δ: 1.03-1.22 (m, 2 H), 1.65 (m, 6 H), 1.86 (m, 1 H), 2.36 (m, 3 H), 2.75(m, 1H), 4.36 (m, 2 H), 6.35 (s, 1H), 7.10-7.27 (m, 6 H), 7.82 (s, 1 H),8.65 (s, 1H). Anal. (C₂₄H₂₅N₇OS• 0.15HOAc•1.2H₂O) C, H, N. APCI-MS: [M +H] 460. Method of Example 31 using 4-chloro-7-methylthieno[3,2-d]pyrimidine in place of 31c.

6,6-dimethyl-N-[(1R,2S)-2-phenylcyclopropyl]-3-(thieno[2,3-d]pyrimidin-4-ylamino)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (CD₃OD) δ:1.01-1.16 (m, 2 H), 1.68 (s, 3 H), 1.69 (s, 3 H), 1.93-2.02 (m, 1 H),2.65-2.74 (m, 1 H), 4.43 (s, 2 H), 7.00-7.09 (m, 3 H), 7.14 (t, J = 7.33Hz, 2 H), 7.34 (d, J = 5.56 Hz, 1 H), 8.03 (d, J = 5.30 Hz, 1 H), 8.58(s, 1 H). LCMS (APCI, M + H⁺): 446.1. Anal.(C₂₃H₂₃N₇OS•1.12H₂O•0.32HOAc): C, H, N. HPLC-UV Detection: 92% purityMethod of Example 32 using 4-chloro-thieno[2,3- d]pyrimidine in place of32c.

6,6-dimethyl-N-[(1R,2S)-2-phenylcyclopropyl]-3-(thieno[3,2-d]pyrimidin-4-ylamino)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (CD₃OD) δ:1.01-1.16 (m, 2 H), 1.68 (s, 3 H), 1.69 (s, 3 H), 1.93-2.02 (m, 1 H),2.66-2.74 (m, 1 H), 4.42 (s, 2 H), 7.00-7.09 (m, 3 H), 7.14 (t, J = 7.33Hz, 2 H), 7.34 (d, J = 5.56 Hz, 1 H), 8.02 (d, J = 5.56 Hz, 1 H), 8.58(s, 1 H). LCMS (APCI, M + H⁺): 446.1. Anal.(C₂₃H₂₃N₇OS•1.08H₂O•0.28HOAc): C, H, N. HPLC: 90% purity. Method ofExample 32 using 4-chlorothieno[3,2- d]pyrimidine in place of 32c.

6,6-dimethyl-3-[(2-methylthieno[3,2-d]pyrimidin-4-yl)amino]-N-[(1R,2S)-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (400 MHz, CH₃OD):δ 1.15-1.19 (m, 2H), 1.79 (s, 3H), 1.80 (s, 3H), 2.01-2.05 (m, 1H),2.76-2.78 (m, 1H), 2.80 (s, 3H), 4.50-4.51 (m, 2H), 7.11-7.15 (m, 3H),7.22-7.25 (m, 2H), 7.46 (d, J = 5.5 Hz, 1H), 8.44 (d, J = 5.3 Hz, 1H).LCMS 460 (M + H). Anal. (C₂₄H₂₅N₇OS•1.50 TFA•0.25 H₂O) C, H, N.HPLC >99% purity. Method of Example 32 using 4-Chloro-2-methylthieno[3,2-d]pyrimidine in place of 32c.

3-(furo[3,2-d]pyrimidin-4-ylamino)-6,6-dimethyl-N-(trans-2-phenylcyclopropyl)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (CD₃OD) δ: 1.01-1.16 (m, 2 H), 1.68(s, 3 H), ¹H NMR (400 MHz, DMSO-d6) δ: 1.03 (m, 1H), 1.22 (m, 1H), 1.64(s, 6H), 1.92 (m, 1H), 2.75 (m, 1H), 4.38 (d, J = 12.1 Hz, 1H), 4.43 (d,J = 12.2 Hz, 1H), 6.35 (s, 1H), 7.05-7.16 (m, 3H), 7.2-7.28 (m, 2H),8.33 (br s, 1H), 8.52 (s, 1H), 10.38 (br s, 1H). Anal.(C₂₃H₂₃N₇O₂•0.2HOAc•1H₂O) C, H, N. APCI- MS: [M + H] 430. Method ofExample 31 using 4-chlorofuro[3,2- d]pyrimidine (prepared according toprocedure reported in WO2004013141, page 131-133) in place of 31c.

Preparation of 40c: 4-Chloroquinazoline-2-carbonitrile

Ethyl 4-quinazolone-2-carboxylate (40a, 2.54 g, 11.67 mmol) wasdissolved in MeOH (29 mL) in a 200 mL round-bottom flask and cooled to0° C. Anhydrous ammonia gas was bubbled into the solution for 30minutes. The flask was then sealed with a suba-seal stopper which wassecured with copper wire. The reaction was then warmed to ambienttemperature and stirred overnight. The solvent was removed under reducedpressure to give 4-Oxo-3,4-dihydroquinazoline-2-carboxamide (40b, 2.20g, 99%) as a white solid. R_(f)=0.13 (7% MeOH/CHCl₃). ¹H NMR (400 MHz,DMSO-d₆): δ 7.58-7.62 (m, 1H), 7.76 (d, J=7.6 Hz, 1H), 7.85-7.89 (m,1H), 8.06 (s, 1H), 8.16 (dd, J=7.8, 1.2 Hz), 8.35 (s, 1H), 11.91 (s,1H). LCMS 190 (M+H). HPLC>99% purity.

4-Oxo-3,4-dihydroquinazoline-2-carboxamide (40b, 0.51 g, 2.69 mmol) washeated to 100° C. in POCl₃ (7.0 mL) for three hours. The solvent wasremoved under reduced pressure. Ice-cold water was carefully added tothe flask, and the insoluble product was filtered off. The filtrate wasextracted three times with chloroform. The combined organics were dried(MgSO₄) and evaporated to give 4-Chloroquinazoline-2-carbonitrile (40c,0.21 g). 95% pure by HPLC and used without further purification. Theinsoluble product was purified by flash silica gel chromatographyeluting with 3-30% EtOAc/hexane to give an additional 0.22 g of 40c as awhite solid for an overall yield of 62%. R_(f)=0.53 (30% EtOAc/hexane).¹H NMR (400 MHz, DMSO-d₆): δ 8.07-8.11 (m, 1H), 8.24-8.31 (m, 2H), 8.41(d, J=8.4 Hz, 1H).

Preparation of 43b: 4-Chloro-5-fluoro-2-methoxypyrimidine

2-Methoxy-5-fluorouracil (43a, 1.04 g, 7.21 mmol) andN,N-dimethylaniline (1.80 mL) were heated in POCl₃ at 110° C. for 90minutes. After cooling, the reaction was added carefully to ice. Theproduct was extracted with diethylether. The ether layer was washed withsequentially with 2N HCl, water, and brine followed by drying (MgSO₄).The ether was carefully removed under reduced pressure to give 43b as avolatile liquid (0.39 g, 34%) which was used without furtherpurification. R_(f)=0.26 (10% EtOAc/hexane). ¹H NMR (400 MHz, DMSO-d₆):δ 3.91 (s, 3H), 8.79 (s, 1H).

Preparation of 44b: 4-Chloro-5-fluoro-2-methylpyrimidine

Sodium hydride (60%, 5.0 g, 125 mmol) was washed with hexane to removethe mineral oil and dried, then suspended in THF (50 mL) and cooled to0° C. Ethyl fluoroacetate (13.30 g, 125 mmol) and ethyl formate (15.14mL, 187 mmol) were mixed together and added to the stirring suspension.The reaction was slowly warmed to ambient temperature and stirred 3days. The solvent was removed. A mixture of acetamidine hydrochloride(11.81 g, 125 mmol), sodium ethoxide (8.86 g, 125 mmol), and ethanol (60mL) were added to the reaction followed by refluxing overnight. Theethanol was removed under reduced pressure. The residue was dissolved ina minimum of water and acidified to pH=6 with concentrated HCl. Thecrude products were then extracted by salting out from the aqueous phaseand washing exhaustively with 4:1 CHCl₃/isopropanol. The combinedorganic phases were dried (MgSO₄) and evaporated. The crude solid waspurified by silica gel chromatography eluting with 5-90% EtOAc/hexane togive 5-Fluoro-2-methylpyrimidin-4(3H)-one (44a, 0.95 g, 6%) as a whitesolid. R_(f)=0.08 (75% EtOAc/hexane). ¹H NMR (400 MHz, DMSO-d₆): δ 2.25(d, J=1.0 Hz, 3H), 7.93 (d, J=3.8 Hz, 1H), 12.95 (br, 1H). LCMS 129.

Compound 44b was prepared following the method of 43b, except that 44awas used in place of 43a. Compound 44b (0.11 g, 10%) was obtained as avolatile liquid and was used without further purification. R_(f)=0.39(5% EtOAc/hexane). ¹H NMR (400 MHz, DMSO-d₆): δ 2.60 (s, 3H), 8.86 (s,1H).

Preparation of Compound 45c:4-chloro-2-(trifluoromethyl)thieno[3,2-d]pyrimidine

To a stirring solution of methyl 3-aminothiophene-2-carboxylate (45a,1.57, 10.0 mmol) in EtOH (10 mL) was added trifluoroacetamidine (2.24 g,2 eq) and trifluoroacetic acid (1.54 ml, 2 eq). The resulting mixturewas heated to 150° C. for 1 hour in microwave reactor. The reactionmixture was cool and filtered to provide2-(trifluoromethyl)thieno[3,2-d]pyrimidine-4(3H)-one 45b as a solid(0.61 g). ¹H NMR (400 MHz, MeOD) δ ppm: 7.49 (d, J=5.29 Hz, 2H), 8.18(d, J=5.29 Hz, 1H).

A suspension of 2-(trifluoromethyl)thieno[3,2-d]pyrimidine-4(3H)-one(45b, 0.61 g, 2.77 mmol) in POCl₃ was refluxed under an atmosphere ofnitrogen for 3 hrs and then concentrated to dryness under reducedpressure. The residue was partitioned between ethyl acetate andsaturated NaHCO₃, dried, and concentrated to give compound 45c as asolid (0.58 g, 88%). ¹H NMR (400 MHz, MeOD) δ ppm: 7.79 (d, J=5.54 Hz,1H), 8.59 (d, J=5.54 Hz, 1H).

Preparation of 48b: 4-Chloro-5-fluoropyrimidine

Sodium hydride (60%, 5.0 g, 125 mmol) was washed with hexane to removethe mineral oil and dried, then suspended in THF (50 mL) and cooled to0° C. Ethyl fluoroacetate (13.35 g, 126 mmol) and ethyl formate (13.99g, 189 mmol) were mixed together and added to the stirring suspension.The reaction was slowly warmed to ambient temperature and stirredovernight. The solvent was removed. A mixture of formamidinehydrochloride (10.33 g, 126 mmol), sodium ethoxide (8.92 g, 126 mmol),and ethanol (60 mL) were added to the reaction followed by refluxingovernight. The ethanol was removed under reduced pressure. The residuewas dissolved in a minimum of water and acidified to pH=6 with ethanolicHCl. The solids were filtered off and the filtrate concentrated. Thecrude solid was purified by silica gel chromatography eluting with 0-9%MeOH/CHCl₃ to give 5-Fluoropyrimidin-4(3H)-one as a white solid (48a,1.05 g, 7%). R_(f)=0.13 (75% EtOAc/hexane). ¹H NMR (400 MHz, DMSO-d₆): δ8.05-8.09 (m, 2H), 13.14 (br, 1H).

Following the procedure to make 43b, compound 48b was synthesized from48a to give 0.97 g (80%) of a volatile liquid which was used withoutfurther purification. R_(f)=0.37 (5% EtOAc/hexane). ¹H NMR (400 MHz,DMSO-d₆): δ 8.93 (s, 1H), 9.01 (s, 1H).

Example 54N-[(1S)-2-(Dimethylamino)-1-phenylethyl]-3-[(6-ethylpyrimidin-4-yl)amino]-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide

Preparation of I(H): 5-tert-butyl 2-ethyl3-amino-6,6-dimethylpyrrolo[3,4-c]pyrazole-2,5(4H,6H)-dicarboxylate

The synthetic route of compound I(H) can be found in Scheme 1 under the“Detailed Description” of this application. The detailed syntheticcondition of for the preparation of I(H) can be found in U.S. PatentApplication Publication No. 2003/0171357 and PCT Publication WO02/12242, the disclosure of which are incorporated herein by reference.

Preparation of Compound 54a: ethyl3-amino-6,6-dimethyl-5,6-dihydropyrrolo[3,4-c]pyrazole-2(4H)-carboxylatedihydrochloride

To a stirred slurry of 5-tert-butyl 1-ethyl3-amino-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-1,5-dicarboxylate(I(H), 30.0 g, 92.5 mmol) in ethanol (200 mL) was drop wise added HCl 4M solution in hexanes (116 mL) drop wise. The resulting clear solutionwas stirred at room temperature for 12 h. The reaction mixture wasconcentrated under vacuum to a residue and stirred with hexane (250 mL)for 10 min. The solid product was collected by filtration, washed withhexane (100 mL) and dried under vacuum at 40° C. for 15 h to get ethyl3-amino-6,6-dimethyl-5,6-dihydropyrrolo[3,4-c]pyrazole-1(4H)-carboxylatedihydrochloride (54a, 27.0 g, 98.5%) as white solid. ¹H NMR (300 MHz,dmso-d₆): δ 1.31 (td, J=7, 1.3 Hz, 3H), 1.59 (s, 6H), 4.09 (t, J=3.7 Hz,2H), 4.36 (qd, J=7.2, 1.2 Hz, 2H), 10.12 (br s, 2H).

Preparation of Compound 54c: benzyl[(1S)-2-(dimethylamino)-2-oxo-1-phenylethyl]-carbamate

To a mixture of (2S)-{[(benzyloxy)carbonyl]amino}(phenyl)acetic acid(54b, 196 g, 688 mmol), HBTU (261 g, 688 mmol), and dichloromethane (2.8L) were added sequentially potassium carbonate (285 g, 2.06 mol) anddimethylamine hydrochloride (84.1 g, 1031 mmol). The reaction mixturewas heated at 40° C. overnight. After cooling to room temperature, thesolids were filtered, washed with ethyl acetate (2×500 mL) and thefiltrate concentrated to a residue. Water (1 L) was added to the residueand the solution kept in an ultrasonic cleanser for 2 hours. Theprecipitated solids were collected and washed with water (4×300 mL),hexane (2×500 mL), and dried under vacuum for 24 hours. The solid crudeproduct was dissolved in chloroform (300 mL) and un-dissolved solidswere filtered off. The filtrate was concentrated to dryness and theresidue dissolved in hexane/ethyl acetate (2:1) (250 mL) and allowed tostand at room temperature overnight. The resulting crystals werecollected by filtration, washed with hexane/ethyl acetate (3:1) (100 mL)and dried in high vacuum at 40° C. for 24 hours to give compound 54c(100.0 g, 47%) as a white crystalline solid. ¹H NMR (CDCl₃) δ: 2.88 (s,3H), 2.98 (s, 3H), 5.01 (d, J=12.2 Hz, 1H), 5.11 (d, J=12.2 Hz, 1H),5.58 (d, J=7.5 Hz, 1H), 6.37 (d, J=7.2 Hz, 1H), 7.32 (m, 10H).

Preparation of Compound 54d: (2S)-2-amino-N,N-dimethyl-2-phenylacetamide

To a solution of 54c (80.0 g, 256 mmol) in ethanol (1.2 L) was added aslurry of Pd/C (10%, 9.0 g) in ethyl acetate (50 mL). The reactionmixture was shaken in Parr-apparatus under hydrogen (40 psi) overnight.The catalyst was removed by filtration through celite. The filter padwas washed with ethanol (2×200 mL) and the combined filtrate wasconcentrated to give 54d (40.2 g, 88%) as a white solid. ¹H NMR (CDCl₃)δ: 2.85 (s, 3H), 2.99 (s, 3H), 4.72 (s, 1H), 7.33 (m, 5H).

Preparation of Compound 54e:N-[(2S)-2-amino-2-phenylethyl]-N,N-dimethylamine

A flask containing dry THF (2300 mL) under a nitrogen atmosphere waschilled by an ice-water bath. Lithium aluminum hydride pellets (59.0 g,1555 mmol) were added. To this LAH suspension, a solution of amide 54d(123.0 g, 691 mmol) in dry THF (800 mL) was slowly added overapproximately 1 hour. The resulting reaction mixture was heated atreflux for 5 hours, then cooled to 10° C. The cooled reaction mixturewas slowly quenched with saturated sodium sulfate solution (380 mL) andstirred overnight. The precipitated solids were filtered off and washedwith ethyl acetate (4×500 mL). The filtrate was concentrated to aresidue that was purified on silica gel column (10% methanol, 5%triethylamine in chloroform) to afford 54e (66.7 g, 59%) as a lightyellow liquid. ¹H NMR (CDCl₃) δ: 2.24 (dd, J=3.6, 12.1 Hz, 1H), 2.29 (s,6H), 2.47 (dd, J=10.6, 12.1 Hz, 1H), 4.07 (dd, J=3.6, 10.4 Hz, 1H), 7.24(m, 1H), 7.37 (m, 4H).

Preparation of Compound 54f:N-[(25)-2-isocyanato-2-phenylethyl]-N,N-dimethylamine hydrochloride

To a cooled (0° C.) and stirred solution of triphosgene (27.1 g, 91.32mmol) in DCM (250 mL) was drop wise added a solution of diisopropylethylamine (23.6 g, 182.26 mmol) in DCM (50 mL) over a period of 20 min. Asolution of N-[(2S)-2-amino-2-phenylethyl]-N,N-dimethylamine (54e, 15.0g, 91.32 mmol) in DCM (100 mL) was drop wise added to the brown reactionmixture while maintaining the temperature below 10° C. The resultingreaction mixture was removed from cooling and stirred for 2 h at roomtemperature. The reaction mixture was concentrated under vacuum to aresidue and stirred with 10% DCM in hexane (50 mL). The solidN-[(2S)-2-isocyanato-2-phenylethyl]-N,N-dimethylamine hydrochloridecompound 54f was separated by filtration and used for the next reactionwithout further purification. (Note: The obtained solid product wasstored under nitrogen). ¹H NMR (300 MHz, dmso-d₆): δ 3.29 (s, 3H), 3.38(s, 3H), 3.68 (t, J=10.1 Hz, 1H), 4.42 (dd, J=11.5, 6.5 Hz, 1H), 5.35(dd, J=9.6, 6.2 Hz, 1H), 7.4-7.6 (m, 5H).

Preparation of Compound 54g: (S)-ethyl3-amino-5-((2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-5,6-dihydropyrrolo[3,4-c]pyrazole-2(4H)-carboxylate

To a cooled (0° C.) and stirred slurry of ethyl3-amino-6,6-dimethyl-5,6-dihydropyrrolo[3,4-c]pyrazole-1(4H)-carboxylatedihydrochloride (54a, 25.0 g, 84.12 mmol) were sequence ally added DIPEA(74 mL, 420.1 mmol) andN-[(2S)-2-isocyanato-2-phenylethyl]-N,N-dimethylamine hydrochloride(54f, 17.1 g, 75.71 mmol). After stirring at room temperature for 10 hunder nitrogen, the mixture was diluted with DCM (100 mL) and washedwith water (2×100 mL). The organic solution was dried (Na₂SO₄),filtered, and concentrated under vacuum. The obtained crude product waspurified on silica gel column (10% MeOH/DCM) to get compound 54g (23.0g, 73.7%) as light yellow solid. M.p: 96-97° C. ¹H NMR (300 MHz,dmso-d₆): δ 1.32 (t, J=7.1 Hz, 3H), 1.51 (s, 3H), 1.57 (s, 3H), 2.19 (s,6H), 2.40 (m, 1H), 2.60 (m, 1H), 4.23 (m, 2H), 4.35 (q, J=6.7 Hz, 2H),4.78 (m, 1H), 6.00 (d, J=6 Hz, 1H), 6.55 (s, 2H), 7.18-7.40 (m, 5H).LCMS (APCI, M+H⁺): 415.

Preparation of Compound 54h:3-Amino-N-[(1S)-2-(dimethylamino)-1-phenylethyl]-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide

Ethyl 3-amino-5-({[(1S)-2-(dimethylamino)-1-phenylethyl]amino}carbonyl)-6,6-dimethyl-5,6-dihydropyrrolo[3,4-c]pyrazole-1(4H)-carboxylate(54g, 9.24 g, 22.30 mmol) was dissolved in MeOH (225 mL). A solution of1N LiOH (36 mL) was added, and the reaction was stirred at ambienttemperature for two hours. The solvent was removed under reducedpressure, the residue diluted with water, and the product was extractedinto 4:1 CHCl₃/iPrOH. The organic layer was separated, washed withbrine, dried (MgSO₄), and evaporated to give compound 54h (7.00 g, 92%)as a yellow amorphous solid which was used without further purification.R_(f)=0.16 (10% methanolic NH₃/CHCl₃). ¹H NMR (400 MHz, CD₃OD): δ 1.60(s, 3H), 1.67 (s, 3H), 2.32 (s, 6H), 2.44 (dd, J=12.9, 4.5 Hz, 1H), 2.78(dd, J=12.6, 10.6 Hz, 1H), 4.34 (d, J=10.3 Hz, 1H), 4.40 (d, J=10.6 Hz,1H), 4.90-4.98 (m, 1H), 7.20-7.24 (m, 1H), 7.28-7.36 (m, 4H). LCMS 343.

Compound 54h (0.18 g, 0.52 mmol) and 4-chloro-6-ethylpyrimidine (54i,0.08 g, 0.574 mmol) were mixed in 1:1 HOAc/H₂O (2.0 mL) and heated to100° C. for 1 hour. The reaction was neutralized with solid NaHCO₃ anddiluted with water and 4:1 CHCl₃/iPrOH. The organic phase was separated,washed with brine, then dried (MgSO₄) and evaporated. The product waspurified by flash silica gel chromatography eluting with 0-5% methanolicNH₃/CHCl₃. The product was further purified by preparative HPLC to givethe titled compound 54N-[(1S)-2-(Dimethylamino)-1-phenylethyl]-3-[(6-ethylpyrimidin-4-yl)amino]-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamideas a white solid (0.077 g, 20%). R_(f)=0.16 (10% methanolic NH₃/CHCl₃).¹H NMR (400 MHz, CD₃OD): δ 1.34 (t, J=7.8 Hz, 3H), 1.72 (s, 3H), 1.78(s, 3H), 2.77-2.82 (m, 2H), 2.97 (s, 3H), 3.05 (s, 3H), 3.45-3.51 (m,1H), 3.63-3.69 (m, 1H), 4.63-4.66 (m, 1H), 4.71-4.76 (m, 1H), 5.43 (dd,J=11.3, 3.8 Hz), 6.94 (br, 1H), 7.32-7.36 (m, 1H), 7.39-7.47 (m, 4H),8.74 (s, 1H). LCMS 449 (M+H). Anal. (C₂₄H₃₂N₈O.2.40TFA.1.0H₂O) C,H,N.HPLC>98% purity.

Example 55N-[(1S)-2-(dimethylamino)-1-phenylethyl]-6,6-dimethyl-3-{[2-(trifluoromethyl)pyrimidin-4-yl]amino}-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide

To 4-chloro-2(trifluoromethyl)pyrimidine (55a, 74 mg, 0.4 mmol),(S)-3-amino-N-(2-(dimethylamino)-1-phenylethyl)-6,6-dimethylpyrrolo[3,4-c]pyrazole-5(1H,4H, 6H)-carboxamide (54h, 141 mg, 0.4 mmol) in IPA (1 mL), was added TEA(114 mL, 0.8 mmol). The reaction was heated in microwave oven at 140° C.for 20 min. HPLC yielded the title compound 55(S)-N-(2-(dimethylamino)-1-phenylethyl)-6,6-dimethyl-3-(2-(trifluoromethylpyrimidin-4-ylamino)pyrrolo[3,4-c]pyrazole-5(1H,4H,6H)-carboxamideas a white powder (8 mg, 4%). ¹H NMR (400 MHz, DCM) δ: 1.30 (m, 6H),1.58 (m, 6H), 3.63 (m, 2H), 3.86 (m, 2H), 4.04 (m, 1H), 7.41-7.50 (m,6H), 8.53 (s, 1H). Anal. (C₂₃H₂₇N₈OF3.2.41TFA.1.7H₂O) C, H, N. APCI-MS:[M+H] 489.

Structure and Example # Chemical name, Analytical data and comments

N-[(1S)-2-(dimethylamino)-1-phenylethyl]-3-[(5-fluoro-6-methylpyrimidin-4-yl)amino]-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. R_(f) = 0.13 (7%methanolic NH₃/CHCl₃). ¹H NMR (400 MHz, CD₃OD): δ 1.67 (s, 3H), 1.74 (s,3H), 2.34 (s, 6H), 2.42 (d, J = 2.8 Hz, 3H), 2.44-2.47 (m, 1H),2.80-2.86 (m, 1H), 4.63-4.68 (m, 2H), 4.97-5.02 (m, 1H), 7.20-7.24 (m,1H), 7.30-7.37 (m, 4H), 8.39 (s, 1H). LCMS 453 (M + H). Anal.(C₂₃H₂₉N₈OF•0.35 H₂O•0.35 hexane) C, H, N. HPLC >98% purity. Method ofExample 54 using 4-Chloro-5-fluoro-6- methylpyrimidine in place of 54i.

N-[(1S)-2-(dimethylamino)-1-phenylethyl]-6,6-dimethyl-3-[(9-methyl-9H-purin-6-yl)amino]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (400 MHz, CD₃OD):δ 1.68 (s, 1H), 1.71 (s, 3H), 2.07 (s, 3H), 2.87 (m, 4H), 3.02 (br s,3H), 3.20 (dd, J = 14.7, 7.3 Hz, 1H), 3.84 (d, J = 14.4 Hz, 1H), 3.96(d, 14.1 Hz, 1H), 4.99 (s, 2H), 7.35-7.7 (m, 5H), 8.03 (td, J = 7.6, 1.1Hz, 1H), 8.20 (d, J = 7.5 Hz, 1H), 8.72 (d, J = 4.6 Hz, 1H). LCMS [M +H]⁺ 475. Anal. (C₂₄H₃₀N₁₀O• 2H₂O•2.74TFA) C, H, N. Method of Example 54using 6-chloro-9-methyl-9H- purine and in place of 54i.

N-[(1S)-2-(dimethylamino)-1-phenylethyl]-6,6-dimethyl-3-[(2-methylthieno[2,3-d]pyrimidin-4-yl)amino]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)- carboxamide ¹H NMR(400 MHz, DMSO-d₆): δ 1.61 (s, 3H), 1.70 (s, 3H), 2.62 (s, 3H), 2.81 (d,J = 4.6 Hz, 3H), 2.91 (d, J = 4.8 Hz, 3H), 3.34 (m, 1H), 3.46 (m, 1H),4.63 (s, 2H), 5.34 (m, 1H), 6.58 (d, J = 9.3 Hz, 1H), 7.18-7.46 (m, 5H),7.58 (d, J = 6 Hz, 1H), 7.83 (br d, J = 5.8 Hz, 1H), 9.13 (br s, 1H),10.39 (s, 1H). LCMS [M + H]⁺ 491 Anal. (C₂₆H₃₀N₈OS•1H₂O•2.55TFA) C, H,N. Method Example 54 using 4-chloro-2- methylthieno[2,3-d]pyrimidine inplace of 54i.

N-[(1S)-2-(dimethylamino)-1-phenylethyl]-3-[(2-ethoxy-5-fluoropyrimidin-4-yl)amino]-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (400 MHz,DMSO-d₆): δ 1.28 (t, J = 7 Hz, 3H), 1.59 (s, 3H), 1.66 (s, 3H), 2.81 (d,J = 4.5 Hz, 3H), 2.88 (d, J = 4.6 Hz, 3H), 4.22 (d, J = 7.1 Hz, 1H),4.26 (d, J = 7 Hz, 1H), 4.49 (s, 2H), 5.34 (m, 1H), 6.59 (d, J = 9.1 Hz,1H), 7.25-7.45 (m, 5H), 8.14 (d, J = 3 Hz, 1H), 8.94 (br s, 2H), 10.03(s, 1H). LCMS [M + H]⁺ 483 Anal. (C₂₄H₃₁N₈FO₂•0.5H₂O•1.64TFA) C, H, N.Method of Example 54 using 4-chloro-2-ethoxy-5- fluoropyrimidine inplace of 54i.

3-[(2-cyanopyrimidin-4-yl)amino]-N-[(1S)-2-(dimethylamino)-1-phenylethyl]-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (CD₃OD) δ: 1.62(s, 3 H), 1.70 (s, 3 H), 2.89 (s, 3 H), 2.97 (s, 3 H), 3.35-3.47 (m, 2H), 4.68-4.88 (m, 2 H), 5.25-5.33 (m, 1 H), 6.83-6.92 (b, 1 H), 7.21-7.41 (m, 5 H), 8.20 (d, J = 7.12 Hz, 1 H). LCMS (APCI, M + H⁺): 446.1.Anal. (C₂₃H₂₇N₉O•1.88TFA•0.15H₂O• 0.04MeCN): C, H, N. HPLC-UV Detection:94% purity Method Example 54 using 4-chloropyrimidine-2- carbonitrile inplace of 54i.

N-[(1S)-2-(dimethylamino)-1-phenylethyl]-6,6-dimethyl-3-(thieno[2,3-d]pyrimidin-4-ylamino)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. R_(f) = 0.18 (7%methanolic NH₃/CHCl₃). ¹H NMR (400 MHz, CD₃OD): δ 1.70 (s, 3H), 1.77 (s,3H), 2.32 (s, 6H), 2.42 (dd, J = 12.9, 4.5 Hz, 1H), 2.78-2.84 (m, 1H),4.63 (d, J = 11.6 Hz, 1H), 4.71 (d, J = 11.3 Hz, 1H), 4.97-5.01 (m, 1H),7.20-7.23 (m, 1H), 7.29-7.36 (m, 4H), 7.43 (d, J = 5.4 Hz, 1H), 8.12 (d,J = 5.3 Hz, 1H), 8.67 (s, 1H). LCMS 477 (M + H). Anal. (C₂₄H₂₈N₈OS•0.40H₂O•0.40 MeOH) C, H, N. HPLC >98% purity. Method of Example 54 using4-Chlorothieno[2,3- cl]pyrimidine in place of 54i.

N-[(1S)-2-(dimethylamino)-1-phenylethyl]-6,6-dimethyl-3-[(6-methylthieno[2,3-d]pyrimidin-4-yl)amino]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)- carboxamide ¹H NMR(CD₃OD) δ: 1.64 (s, 3 H), 1.70 (s, 3 H), 2.54 (s, 3 H), 2.89 (s, 3 H),2.97 (s, 3 H), 3.37-3.46 (m, 1 H), 3.52 (t, J = 11.37 Hz, 1 H), 4.57 (d,J = 11.62 Hz, 1 H), 4.66 (d, J = 11.37 Hz, 1 H), 5.33 (dd, J = 4.04,11.37 Hz, 1 H), 7.22-7.37 (m, 6 H), 8.42 (s, 1 H). LCMS (APCI, M + H⁺):491.3. Anal. (C₂₅H₃₀N₈OS• 2.00TFA•0.77H₂O): C, H, N. HPLC 94% purityMethod of Example 54 using 4-chloro-6- methylthieno[2,3-d]pyrimidine inplace of 54i.

N-[(1S)-2-(dimethylamino)-1-phenylethyl]-6,6-dimethyl-3-[(2-methylthieno[3,2-d]pyrimidin-4-yl)amino]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)- carboxamide R_(f) =0.24 (7% methanolic NH₃/CHCl₃). ¹H NMR (400 MHz, CD₃OD): δ 1.77 (s, 3H),1.80 (s, 3H), 2.77 (s, 3H), 2.94 (s, 3H), 3.05 (s, 3H), 3.42-3.46 (m,1H), 3.63-3.69 (m, 1H), 4.66 (s, 2H), 5.39 (dd, J = 11.4, 4.0 Hz),7.33-7.47 (m, 6H), 8.42 (d, J = 5.3 Hz, 1H). LCMS 491 (M + H). Anal.(C₂₅H₃₀N₈OS•3.0 TFA•0.40 H₂O) C, H, N. Method of Example 54 using4-Chloro-2- methylthieno[3,2-d]pyrimidine in place of 54i.

3-[(2-cyanoquinazolin-4-yl)amino]-N-[(1S)-2-(dimethylamino)-1-phenylethyl]-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. R_(f) = 0.29 (7%methanolic NH₃/CHCl₃). ¹H NMR (400 MHz, CH₃OD): δ 1.71 (s, 3H), 1.77 (s,3H), 2.35 (s, 6H), 2.36-2.37 (m, 1H), 2.79-2.85 (m, 1H), 4.91- 5.00 (m,3H), 7.21-7.24 (m, 1H), 7.31-7.35 (m, 2H), 7.38-7.40 (m, 2H), 7.76-7.80(m, 1H), 7.91-7.92 (m, 1H), 7.95-7.99 (m, 1H), 8.42-8.43 (m, 1H). LCMS496 (M + H). Anal. (C₂₇H₂₉N₉O•0.60 H₂O•0.10 hexane) C, H, N. Method ofExample 54 using 4-Chloroquinazoline-2- carbonitrile in place of 54i.

N-[(1S)-2-(dimethylamino)-1-phenylethyl]-6,6-dimethyl-3-[(2-methylpyrimidin-4-yl)amino]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (400 MHz, DMSO)δ: 1.58 (m, 6 H), 2.42 (m, 6H), 4.65 (m, 2 H), 5.13 (m, 1H), 6.64 (m,1H), 7.23- 7.41 (m, 6 H), 8,14 (d, J = 3 Hz), 1H. APCI-MS: [M + H] 435.Method of Example 54 using 4-chloro-2- methylpyrimidine in place of 54i.

N-[(1S)-2-(dimethylamino)-1-phenylethyl]-3-[(5-fluoro-2-methoxypyrimidin-4-yl)amino]-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. ¹H NMR (400 MHz,CD₃OD): δ 1.72 (s, 3H) 1.77 (s, 3H), 3.06 (s, 3H), 3.12 (s, 3H),3.46-3.50 (m, 1H), 3.56-3.62 (m, 1H), 3.89 (s, 3H), 4.63-4.66 (m, 2H),5.37-5.41 (m, 1H), 7.35-7.44 (m, 5H), 8.07 (d, J = 3.3 Hz, 1H). LCMS 469(M + H). Anal. (C₂₃H₂₉N₈OF•2.25 TFA) C, H, N. Method of Example 54 using4-Chloro-5-fluoro-2- methoxypyrimidine in place of 54i.

N-[(1S)-2-(dimethylamino)-1-phenylethyl]-3-[(5-fluoropyrimidin-4-yl)amino]-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (400 MHz, CD₃OD):δ 1.72 (s, 3H) 1.78 (s, 3H), 2.98 (s, 3H), 3.06 (s, 3H), 3.47-3.54 (m,1H), 3.58-3.64 (m, 1H), 4.65 (d, J = 11.4 Hz, 1H), 4.74 (d, J = 11.4 Hz,1H, 5.42 (dd, J = 11.4, 4.0 Hz, 1H), 7.33- 7.36 (m, 1H), 7.39-7.44 (m,4H), 8.32 (s, 1), 8.52 (s, 1H). LCMS 439 (M + H). Anal. (C₂₂H₂₇N₈OF•2.20TFA•0.20 H₂O) C, H, N. HPLC = 91% purity Method of Example 54 using4-Chloro-5- fluoropyrimidine in place of 54i.

Example 68 (1S)-2-(dimethylamino)-1-phenylethyl6,6-dimethyl-3-(thieno[3,2-d]pyrimidin-4-ylamino)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate

Preparation of Compound 68b: (S)-2-dimethylamino-1-phenyl-ethanol

To a solution of (S)-(+)-2-amino-1-phenyl-ethanol (68a, 100.0 g, 729.0mmol) in formic acid (400 mL) was added formaldehyde (800 mL, 37% wt inwater) at room temperature. The solution was stirred at 95° C.overnight. After it was cooled to room temperature, conc. HCl was usedto adjust the solution to pH=2. It was extracted with ether (3×500 mL)and then adjusted to pH=10 with solid NaOH. The resulting aqueous layerwas extracted with CH₂Cl₂ (3×500 mL). The combined organic layers weredried over Na₂SO₄. Filtration and evaporation followed by flashchromatography (5% MeOH in CH₂Cl₂ to 4.5% MeOH/0.5% NEt₃ in CH₂Cl₂) gavecompound 68b (S)-2-dimethylamino-1-phenyl-ethanol as a light-yellow oil(68.0 g, 56%). ¹H NMR (300 MHz, CDCl₃) δ: 2.35 (s, 6H), 2.37 (m, 1H),2.46 (dd, J=12.8, 9.2 Hz, 1H), 4.02 (br s, 1H), 4.69 (dd, J=10.5, 3.6Hz, 1H), 7.22-7.4 (m, 5H).

Preparation of Compound 68e: (S)-5-(2-(dimethylamino)-1-phenylethyl)2-ethyl3-amino-6,6-dimethylpyrrolo[3,4-c]pyrazole-2,5(4H,6H)-dicarboxylate

To a stirred solution of (S)-2-dimethylamino-1-phenyl-ethanol (68b,21.50 g, 130.0 mmol) in 1,2-dichloroethane (500 mL) was addedtriethylamine (26.30 g, 260.0 mmol) and 4-nitrophenyl chloroformate(68c, 27.00 g, 130.0 mmol) at room temperature under nitrogen. Thesolution was stirred at 50° C. overnight. A total of 16.8 g (130.0 mmol)of Hünig's base was then added, followed by ethyl3-amino-6,6-dimethyl-5,6-dihydropyrrolo[3,4-c]pyrazole-1(4H)-carboxylatedihydrochloride salt (54a, 17.90 g, 60.25 mmol). The reaction mixturewas stirred at 50° C. for another 12 h. It was diluted withdichloromethane (1.5 L) and washed with water (2×1.0 L) and brine (1.0L), dried over Na₂SO₄. Another batch with the exact scale was alsocarried out. These two batches were combined together during workup.Filtration and evaporation followed by flash chromatography (4.75%MeOH/0.25% NEt₃/95% DCM) afforded compound 68e ethyl3-amino-5({[(1S)-2-(dimethylamino)-1-phenylethyl]hydroxy}carbonyl)-6,6-dimethyl-5,6-dihydropyrrolo[3,4-c]pyrazole-1(4H)-carboxylateas a light-yellow gummy oil (5.00 g, 10%). ¹H NMR (CDCl₃, a mixture ofrotamers, only the chemical shifts of the major form is reported) δ:1.45 (t, J=7.1 Hz, 3H), 1.63 (s, 3H), 1.72 (s, 3H), 2.29 (s, 3H), 2.36(s, 3H), 2.55-2.63 (m, 1H), 2.88 (dd, J=13, 8.3 Hz, 1H), 4.29 (q, J=13Hz, 1H), 4.51 (q, J=7.1 Hz, 2H), 5.44 (d, J=10.7 Hz, 1H), 5.8-5.95 (m,1H), 7.25-7.42 (m, 5H). LCMS (APCI, M+H⁺) 416.

Preparation of Compound 68f: (S)-2-(dimethylamino)-1-phenylethyl3-amino-6,6-dimethylpyrrolo[3,4-c]pyrazole-5(1H,4H,6H)-carboxylate

A round bottom flask was charged with compound 68e (1.01 g, 0.242 mmol),1N LiOH (3.87 ml, 1.6 eq) and methanol (24 ml). The resulting mixturewas stirred at room temperature for 3 hours. Solvent was evaporated. Tothe residue were added ethyl acetate (20 ml) and water (20 ml). Thewater phase was separated and extracted with ethyl acetate (10 ml). Thecombined ethyl acetate phase was washed with brine, dried with anhydrousNa₂SO₄, filtered and evaporated to give the crude compound 68f (564 mg,67%). LCMS (APCI, M+H⁺): 344.1.

A re-sealable tube was charged with compound 68f (88 mg, 0.257 mmol),4-chloro-thieno (3,2-d)-pyrimidine (88 mg, 2 eq), and a mixture ofacetic acid and water (1 to 1, 1 ml). The tube was capped and stirred at100° C. for 1 hour. It was then purified twice by prep. HPLC to givetitle compound 68 as a white solid (36 mg, 29% yield). ¹H NMR (CD₃OD, amixture of rotamers, only the chemical shifts of the major form isreported) δ: 1.55 (s, 3H), 1.66 (s, 3H), 2.62 (s, 6H), 2.82-2.92 (m,1H), 3.21-3.37 (m, 1H), 4.44-4.57 (m, 1H), 4.60-4.69 (m, 1H), 5.90-6.00(m, 1H), 7.22-7.43 (m, 6H), 8.01-8.09 (m, 1H), 8.55 (s, 1H). LCMS (APCI,M+H⁺): 478.2. Anal. (C₂₄H₂₇N₇O₂S.0.2HOAc.0.45TFA.0.98H₂O): C, H, N.

Structure and Example # Chemical name, Analytical data and comments

(1S)-2-(dimethylamino)-1-phenylethyl 6,6-dimethyl-3-[(2-methylthieno[2,3-d]pyrimidin-4-yl)amino]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate ¹H NMR (400 MHz,DMSO-d₆) δ: 1.55 (s, 3H), 1.67 (s, 3H), 2.63 (s, 3H), 2.87 (d, J = 4.8Hz, 3H), 2.93 (d, J = 5 Hz, 3H), 3.46 (m, 1H), 3.60 (m, 1H), 3.84 (d, J= 13.1 Hz, 1H), 3.97 (d, J = 13.1 Hz, 1H), 6.13 (dd, J = 10.7, 2.3 Hz,1H), 7.35-7.50 (m, 5H), 7.58 (d, J = 6.1 Hz, 1H), 7.87 (d, j = 6 Hz,1H), 9.49 (br s, 1H), 10.45 (s, 1H), LCMS [M + H]⁺ 492 Anal(C₂₅H₂₉N₇O₂S•1.8H₂O•0.44TFA•0.4HOAc) C, H, N, S. Method of Example 68using 4-chloro-2-methylthieno[2,3- d]pyrimidine in place of 68g.

(1S)-2-(dimethylamino)-1-phenylethyl 3-[(2-cyanopyrimidin-4-yl)amino]-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate ¹H NMR (CD₃OD) δ: 1.49 (s, 3 H), 1.64(s, 3 H), 2.93 (s, 3 H), 2.97 (s, 3 H), 3.30-3.39 (m, 1 H), 3.59-3.70(m, 1 H), 4.92 (d, J = 13.39 Hz, 1 H), 5.14 (d, J = 13.39 Hz, 1 H), 6.03(dd, J = 2.27, 10.86 Hz, 1 H), 6.86 (d, J = 6.06 Hz, 1 H), 6.86 (d, J =6.06 Hz, 1 H), 7.25-7.44 (m, 5 H), 8.19- 8.25 (m, 1 H). LCMS (APCI, M +H⁺): 447.1. Anal. (C23H26N8O₂•1.91TFA): C, H, N. HPLC-UV Detection: >95%purity. Method of Example 68 using 4-chloropyrimidine-2- carbonitrile inplace of 68g.

(1S)-2-(dimethylamino)-1-phenylethyl 6,6-dimethyl-3-[(6-methylthieno[2,3-d]pyrimidin-4-yl)amino]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate ¹H NMR (CD₃OD, a mixtureof rotamers, only the chemical shifts of the major form is reported) δ:1.55 (s, 3 H), 1.66 (s, 3 H), 2.55 (s, 3 H), 2.92 (s, 3 H), 2.99 (s, 3H), 3.33-3.46 (m, 1 H), 3.64-3.81 (m, 1 H), 4.63-4.80 (m, 2 H),6.04-6.14 (m, 1 H), 7.22-7.45 (m, 6 H), 8.41 (s, 1 H). LCMS (APCI, M +H⁺): 492.3. Anal. (C₂₅H₂₉N₇O₂S• 1.74TFA•0.58H₂O): C, H, N. HPLC-UVDetection: 93% purity. Method of Example 68 using4-chloro-6-methylthieno [2,3-d]pyrimidine in place of 68g.

(1S)-2-(dimethylamino)-1-phenylethyl 3-[(2-ethoxy-5-fluoropyrimidin-4-yl)amino]-6,6-dimethyl-4,6-dihydro-pyrrolo[3,4-c]pyrazole-5(1H)-carboxylate. ¹H NMR (CD₃OD, a mixture ofrotamers, only the chemical shifts of the major form is reported) δ:1.25 (t, J = 7.07 Hz, 3 H), 1.54 (s, 3 H), 1.65 (s, 3 H), 2.91 (s, 3 H),2.98 (s, 3 H), 3.36-3.44 (m, 1 H), 3.58-3.69 (m, 1 H), 4.25 (q, J = 7.07Hz, 2 H), 4.70 (s, 2 H), 6.07-6.15 (m, 1 H), 7.28-7.43 (m, 5 H), 7.97(d, J = 3.28, 1 H). LCMS (APCI, M + H⁺): 484.2. Anal.(C₂₄H₃₀FN₇O₃•2.06TFA•0.31H₂O): C, H, N; HPLC-UV Detection: 100% purity.Method of Example 68 using 4-chloro-2-ethoxy-5- fluoropyrimidine inplace of 68g.

(1S)-2-(dimethylamino)-1-phenylethyl 6,6-dimethyl-3-[(2-methylthieno[3,2-d]pyrimidin-4-yl)amino]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate ¹H NMR (400 MHz, CD₃OD):δ 1.67 (s, 3H), 1.78 (s, 3H), 2.79 (s, 3H), 3.07 (s, 6H), 3.48 (dd, J =13.9, 2.5 Hz, 1H), 3.80 (dd, J = 13.6, 10.6 Hz, 1H), 4.71-4.99 (m, 2H),6.21- 6.22 (m, 1H), 7.39-7.58 (m, 6H), 8.42 (d, J = 5.5 Hz, 1H). LCMS492 (M + H). Anal. (C₂₅H₂₉N₇O₂S•2.75 TFA) C, H, N. Method of Example 73using 4-Chloro-2- methylthieno[3,2-d]pyrimidine in place of 68g.

(1S)-2-(dimethylamino)-1-phenylethyl 3-[(2-cyanoquinazolin-4-yl)amino]-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate. ¹H NMR (400 MHz,CD₃OD): δ 1.62 (s, 3H), 1.77 (s, 3H), 3.03 (s, 3H), 3.07 (s, 3H),3.42-3.46 (m, 1H), 3.76-3.82 (m, 1H), 5.13 (d, J = 13.3 Hz, 1H), 5.36(d, J = 13.3 Hz, 1H), 6.14 (dd, J = 11.1, 2.3 Hz, 1H) 7.36-7.46 (m, 3H),7.52-7.54 (m, 2H), 7.78-7.82 (m, 1H), 7.92 (d, J = 8.3 Hz, 1H),7.97-8.01 (m, 1H), 8.45 (d, J = 8.3 Hz, 1H). LCMS 497 (M + H). Anal.(C₂₇H₂₈N₈O₂•1.9 TFA•0.25 water) C, H, N. Method of Example 68 using4-Chloroquinazoline-2- carbonitrile in place of 68g.

(1S)-2-(dimethylamino)-1-phenylethyl 6,6-dimethyl-3-(thieno[2,3-d]pyrimidin-4-ylamino)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate. R_(f) = 0.26 (7% methanolic NH₃/CHCl₃). ¹HNMR (400 MHz, CD₃CN): δ 1.59 (s, 3H), 1.68 (s, 3H), 2.28 (s, 6H),2.51-2.58 (m, 1H), 2.77-2.82 (m, 1H), 4.70 (s, 2H), 5.80-5.86 (m, 1H),7.27-7.46 (m, 5H), 8.01-8.04 (m, 1H), 8.58 (br, 1H), 8.68 (d, J = 8.5Hz, 1H). LCMS 478 (M + H). Anal. (C₂₄H₂₇N₇O₂S•0.06 TFA•0.10 hexane) C,H, N. HPLC >99% purity. Method of Example 68 using 4-Chlorothieno[2,3-d]pyrimidine in place of 68g.

(1S)-2-(dimethylamino)-1-phenylethyl 6,6-dimethyl-3-{[2-(trifluoromethyl)pyrimidin-4-yl]amino}-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate ¹H NMR (400 MHz, DMSO)δ: 1.58 (m, 6 H), 2.22 (m, 6H), 4.76 (m, 1 H), 5.79 (m, 1H), 7.29-7.40(m, 5 H). APCI-MS: [M + H] 490. Method of Example 68 using 4-chloro-2-(trifluoromethyl)pyrimidine in place of 68g.

(1S)-2-(dimethylamino)-1-phenylethyl 3-[(5-fluoro-6-methylpyrimidin-4-yl)amino]-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate ¹H NMR (CD₃OD, a mixtureof rotamers, only the chemical shifts of the major form is reported) δ:1.52 (s, 3 H), 1.63 (s, 3 H), 2.28-2.36 (m, 1 H), 2.34 (s, 3 H), 2.39(s, 3 H), 2.52 (s, 3 H), 2.68-2.83 (m, 1 H), 4.43-4.65 (m, 2 H),5.84-5.95 (m, 1 H), 7.18-7.41 (m, 5 H), 8.25 (s, 1 H). LCMS (APCI, M +H⁺): 454.2. Anal. (C₂₃H₂₈FN₇O₂• 0.45TFA•0.40HOAc•0.20H₂O) C, H, N.HPLC-UV Detection: 100% purity. Method of Example 68 using4-chloro-5-fluoro-6- methylpyrimidine in place of 68g.

(1S)-2-(dimethylamino)-1-phenylethyl 6,6-dimethyl-3-[(9-methyl-9H-purin-6-yl)amino]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate ¹H NMR (CD₃OD, a mixture of rotamers, onlythe chemical shifts of the major form is reported) δ: 1.55 (s, 3 H),1.67 (s, 3 H), 2.94 (s, 3 H), 3.00 (s, 3 H), 3.36-3.43 (m, 1 H),3.69-3.78 (m, 1 H), 3.81 (s, 1 H), 4.72-4.81 (m, 2 H), 6.07-6.12 (m, 1H), 7.29-7.44 (m, 5 H), 8.14 (s, 1 H), 8.44 (s, 1 H). LCMS (APCI, M +H⁺): 476.2. Anal. (C₂₄H₂₉N₉O₂•1.87TFA•0.73H₂O): C, H, N. HPLC-UVDetection: 100% purity. Method of Example 78 using 6-chloro-9-methyl-9H-purine in place of 68g.

(1S)-2-(dimethylamino)-1-phenylethyl 3-[(5-fluoro-2-methoxypyrimidin-4-yl)amino]-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate. ¹H NMR (400 MHz,CD₃OD): δ 1.62 (s, 3H), 1.73 (s, 3H), 3.02 (s, 3H), 3.09 (s, 3H),3.47-3.50 (m, 1H), 3.72-3.78 (m, 1H), 3.93 (s, 3H), 4.58-4.61 (m, 2H),6.18-6.20 (m, 1H), 7.41-7.57 (m, 5H), 8.07 (d, J = 3.0 Hz, 1H). LCMS 470(M + H). Anal. (C₂₃H₂₈N₇O₃F•2.30 TFA•0.25 H₂O) C, H, N. HPLC >99%purity. Method of Example 68 using 4-Chloro-5-fluoro-2-methoxypyrimidine in place of 68g.

(1S)-2-(dimethylamino)-1-phenylethyl 3-[(5-fluoropyrimidin-4-yl)amino]-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate. ¹H NMR (400 MHz,CH₃OD): δ 1.61 (s, 3H), 1.71 (s, 3H), 2.35 (s, 3H), 2.42 (s, 3H),2.55-2.65 (m, 1H), 3.08-3.11 (m, 1H), 4.93-4.98 (m, 2H), 5.87-5.96 (m,1H), 7.28-7.45 (m, 5H), 8.28 (br, 1H), 8.48 (br, 1H). LCMS 440 (M + H).Anal. (C₂₂H₂₆N₇O₂F•0.70 H₂O•0.30 hexane) C, H, N. PLC = 90% purity.Method of Example 68 using 4-Chloro-5- fluoropyrimidine in place of 68g.

Example 81N-(6,6-dimethyl-5-{[(3S,8aS)-3-methylhexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl]carbonyl}-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-7-methylthieno[3,2-d]pyrimidin-4-amine

Preparation of compound 81b:(3S,8aS)-3-methylhexahydropyrrolo[1,2-a]pyrazine-2(1H)-carbonyl chloride

To a stirring mixture of triphosgene (2.11 g, 1 eq) in CH₂Cl₂ (10 ml) at0° C. was added DIPEA (1.8 ml, 1.5 eq) and(3S,8aS)-3-methyloctahydropyrrolo[1,2-a]pyrazine (81a, 1 g, 7.13 mmol).The resulting mixture was stirred at 0° C. for 30 min. The reactionmixture was evaporated in vacuo to give a residue, compound 81b, whichwas directly carried onto the next reaction without furtherpurification.

Preparation of Compound 81c: ethyl3-amino-6,6-dimethyl-5-{[(3S,8aS)-3-methylhexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl]carbonyl}-5,6-dihydropyrrolo[3,4-c]pyrazole-2(4H)-carboxylate

To a stirring mixture of compound I(H) 5-tert-butyl 2-ethyl3-amino-6,6-dimethylpyrrolo[3,4-c]pyrazole-2,5(4H,6H)-dicarboxylate(5.65 g, 17.4 mmol) in CH₂Cl₂ (20 ml) was added 4.0M HCl in dioxane (30ml). The reaction mixture was concentrated in vacuo to give crude HClsalt of compound 54a. A portion of residue (54a, 1 g, 4.46 mmol) wasadded to a stirring mixture of(3S,8aS)-3-methylhexahydropyrrolo[1,2-a]pyrazine-2(1H)-carbonyl chloride(81b, 1.4 g, 2 eq) in CH₂Cl₂ (20 ml), DIPEA (1.2 ml, 2 eq). Theresulting mixture was stirred at room temperature for 15 h. The reactionmixture was diluted with CH₂Cl₂, and washed with saturated NaHCO₃, driedover sodium sulfate, concentrated in vacuo, purified by flashchromatography. Elution with 5-15% MeOH/DCM provided compound 81c. ¹HNMR (CD₃)₂SO δ: 1.2 (m, 2H), 1.31 (t, 3H), 1.52 (m, 6H), 1.64 (m, 4H),1.93 (m, 1H), 2.18 (m, 1H), 2.77 (m, 2H), 2.93 (m, 1H), 3.77 (m, 1H),4.18 (m, 2H), 4.33 (m, 2H)

Preparation of Compound 81d:6,6-dimethyl-5-{[(3S,8aS)-3-methylhexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl]carbonyl}-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-amine

To a stirring solution of ethyl3-amino-6,6-dimethyl-5-{[(3S,8aS)-3-methylhexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl]carbonyl}-5,6-dihydropyrrolo[3,4-c]pyrazole-2(4H)—carboxylate (81c, 613 mg, 1.60 mmol) in MeOH (3 mL) was added 20% aq.NaOH (2 ml). The resulting mixture was stirred at room temperature for30 min. The reaction mixture was concentrated and the residue waspartitioned between ethyl acetate and saturated NaHCO₃, dried, andconcentrated to give compound 81d.

To a stirring solution of compound 81d (0.150 g, 0.47 mmol) in 50%acetic acid/water (4 ml) was added4-chloro-7-methylthieno[3,2-d]pyrimidine (175 mg, 2 eq). The resultingmixture was heated to a temperature of 100° C. for 1 hr. The reactionmixture was purified by prep-HPLC to provide compound 81 as a whitesolid ¹H NMR (CD₃)₂SO δ: 1.23 (m, 2H), 1.62 (d, 6H), 1.69 (m, 3H), 1.83(m, 1H), 1.95 (m, 1H), 2.16 (m, 1H), 2.34 (s, 3H), 2.74 (m, 2H), 2.90(m, 1H), 3.80 (m, 1H), 4.52 (s, 2H), 7.83 (s, 1H), 8.56 (s, 1H).

Structure and Example # Chemical name, Analytical data and comments

N-(5-fluoro-6-methylpyrimidin-4-yl)-6,6-dimethyl-5-{[(3S,8aS)-3-methylhexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl]carbonyl}-1,4,5,6-tetrahydropyrrolo[3,4- c]pyrazol-3-amine ¹HNMR (400 MHz, (CD₃)₂SO δ: 1.23 (m, 2 H), 1.27 (m, 1H), 1.59 (d, 6 H),1.70 (m, 2 H), 1.83 (m, 1H), 1.94 (m, 1H), 2.16 (m, 1H), 2.32 (s, 3H),2.70 (m, 2H), 2.90 (t, 1H), 3.79 (m, 1H), 4.48 (s, 2H), 8.23 (s, 1H).Method of Example 81 using 4-chloro-5-fluoro-6- methylpyrimidine wasused in place of 81e.

More Examples

Structure and Example # Chemical name, Analytical data and comments

N-[(1S)-2-(dimethylamino)-1-phenylethyl]-3-(furo[3,2-d]pyrimidin-4-ylamino)-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide ¹H NMR (400 MHz, CD₃OD):δ 1.74 (s, 3H) 1.80 (s, 3H), 2.98 (s, 3H), 3.06 (s, 3H), 3.47-3.53 (m,1H), 3.61-3.67 (m, 1H), 4.70 (d, J = 11.6 Hz, 1H), 4.77 (d, J = 11.3 Hz,1H), 5.41-5.45 (m, 1H), 7.06 (d, J = 2.2 Hz, 1H), 7.32-7.36 (m, 1H),7.40-7.46 (m, 4H), 8.25 (d, J = 2.3 Hz, 1H), 8.67 (d, J = 3.0 Hz, 1H).LCMS 461 (M + H). Anal. (C₂₄H₂₈N₈O₂•2.40 TFA•0.40 H₂O) C, H, N. HPLC =95% purity Method of Example 54 using 4-chlorofuro[3,2- d]pyrimidine(prepared according to procedure reported in WO 2004013141 page 131-133)in place of 54i.

3-[(2-methylquinazolin-4-yl)amino]-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. ¹H NMR (400 MHz, MeOD) δ ppm: 1.12-1.25(m, 2 H), 1.79 (d, J = 3.78 Hz, 6 H), 2.01-2.09 (m, 1 H), 2.67 (s, 3 H),2.77-2.86 (m, 1 H), 4.55 (s, 2 H), 7.09- 7.18 (m, 3 H), 7.24 (t, J =7.55 Hz, 2 H), 7.57 (t, J = 7.68 Hz, 1 H), 7.72 (d, J = 8.06 Hz, 1 H),7.83 (t, J = 7.55 Hz, 1 H), 8.30 (d, J = 8.31 Hz, 1 H). Anal.(C₂₆H₂₇N₇O•0.3HOAc•0.5H₂O) C, H, N. HPLC: >95% purity. Method of Example31 using 4-chloro-2- methylquinazoline in place of 31c.

3-(quinazolin-4-ylamino)-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4- c]pyrazole-5(1H)-carboxamide.¹H NMR (400 MHz, MeOD) δ ppm: 1.12-1.27 (m, 2 H), 1.79 (d, J = 3.53 Hz,6 H), 2.03-2.16 (m, 1 H), 2.81 (dd, J = 6.42, 3.15 Hz, 1 H), 4.58 (s, 2H), 7.08- 7.19 (m, 3 H), 7.24 (t, J = 7.68 Hz, 2 H), 7.65 (t, J = 7.55Hz, 1 H), 7.77-7.96 (m, 2 H), 8.35 (d, J = 8.31 Hz, 1 H), 8.71 (s, 1 H).Anal. (C₂₅H₂₅N₇O•0.3HOAc•0.6H₂O) C, H, N. HPLC: >95% purity. Method ofExample 31 using 4-chloroquinazoline in place of 31c.

3-[(2-cyclopropylquinazolin-4-yl)amino]-6,6-dimethyl-N-[trans-2-phenylcyclopropyl]-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. ¹H NMR (400 MHz, MeOD) δ ppm: 1.00-1.11(m, J = 6.55 Hz, 2 H), 1.11-1.24 (m, 4 H), 1.79 (d, J = 3.53 Hz, 6 H),2.00-2.09 (m, 1 H), 2.14-2.27 (m, 1 H), 2.73-2.87 (m, 1 H), 4.44 (s, 2H), 7.07-7.18 (m, 3 H), 7.24 (t, J = 7.68 Hz, 2 H), 7.52 (t, J = 7.43Hz, 1 H), 7.67-7.76 (m, 1 H), 7.81 (t, J = 7.55 Hz, 1 H), 8.25 (d, J =8.06 Hz, 1 H). Anal. (C₂₈H₂₉N₇O•0.2HOAc•0.6H₂O) C, H, N. HPLC: >95%purity. Method of Example 31 using 4-chloro-2- cyclopropylquinazoline inplace of 31c.

Biological Testing, Ki Data and Cellular Assay Data

Cloning, expression, and purification of recombinant PAK4 Kinase domain(PAK4 KD): The cDNA coding for PAK4 was amplified from the EST clone(#12) (purchased from Research Genetics) by using PCR. P33 (ACATATG TCCCATGAGCAGT TCCGGGCTGC CCTGCAGCT) and P34 (CTCA TGGGTGCTTC AGCAGCTCGGCTGCCGTGGC) were used as the 5′ primer and 3′ primer in PCRrespectively. The PCR amplified product was cloned into Topo vector(Invitrogen Inc.), and verified by DNA sequencing. PAK4 KD was thensubcloned into expression plasmid pET28a(+), pET24a(+), or pGST4.5. Therecombinant plasmids containing PAK4 KD was transformed into BL21(DE3)cells for recombinant protein expression. The production of PAK4 KD wasinduced at 27° C. by the addition of IPTG into the cells. The cells werethen harvested and lyzed for protein purification. Ni-NTA column(pET28a(+), pET24a(+)) and glutathione column (pGST4.5) were used forthe purification. The purified protein was then subjected to thrombin tocleave the N-terminal tags that were inherited from the expressionplasmids, and thus gave the PAK4 KD that were used for the Ki assay ofthis invention.

PAK4 kinase domain enzymatic assay conditions: the enzymatic activity ofPAK4 KD was measured by its ability to catalyze the transfer of aphosphate residue from a nucleoside triphosphate to an amino acid sidechain of a commercially available peptide (amino acid sequenceEVPRRKSLVGTPYWM). The conversion of ATP to ADP accompanies the catalyticreaction. The PAK4 KD catalyzed production of ADP from ATP was coupledto the oxidation of NADH through the activities of pyruvate kinase (PK)and lactate dehydrogenase (LDH). The conversion of NADH to NAD⁺ ismonitored by the decrease in absorbance at 340 nm (e340=6.22 cm⁻¹mM⁻¹)using a Molecular Devices SPECTRAMAX 190 in conjunction with the BiomecFX. Typical reaction solutions contain 2 mM phosphoenolpyruvate, 0.35 mMNADH, 10 mM MgCl₂, 1 mM DTT, 0.4 mM peptide (EVPRRKSLVGTPYWM) 0.04 mMATP, 1 units/mL PK, 1 units/mL LDH, 0.01% Tween 20 in 50 mM HEPES, pH7.5. Assays are initiated with the addition of 25 nM PAK4 KD. The PAK KDKi of each compound of the invention (the inhibitor) was calculatedbased on multiple of Percent Inhibition numbers of the inhibitor atdifferent inhibitor concentrations. The peptide (amino sequenceEVPRRKSLVGTPYWM) was purchased from American Peptide Company. NADH,MgCl₂, HEPES, DTT, ATP and PK/LDH were purchased from Sigma. Tween 20was purchased from Calbiochem.

A sandwich ELISA method was used to measure the PAK4 kinase activity inwhole cells. The level of PAK4-dependent phosphorylation of GEF-H1b canbe determined by monitoring the binding of a phosphospecific antibody toGEF-H1b. A modified HEK 293 cell line is used in the bioassay and it hasbeen engineered to overexpress both GEF-H1b and the kinase domain (KD)of PAK4. The KD of PAK4 is inducible in this cell line by tetracycline(Trex system, Invitrogen). The name of this cell line has beendesignated TR-293-KDG. To establish a phosphorylation event on GEF-H1,cells are induced with doxycycline to express the PAK4 KD. Negativecontrol wells do not receive induction. Candidate substance effect ismeasured as the ability to block this phosphorylation event.

ELISA plate was prepared by pre-coating the plates with a captureantibody (α-HA-tag mouse monoclonal antibody), blocked with BSA, andwashed in 0.1% tween 20 in tris-buffered saline (TBST). Tissue cultureplates (precoated with poly-D-lysine) were seeded with TR-293-KDG cells.The TR-293-KDG Cells were induced to express the PAK4 KD withdoxycycline overnight and subsequently & concomitantly treated withcandidate substances or diluent for an additional 3-hour, continuousexposure. Cells were then lysed with a modified RIPA buffer supplementedwith protease inhibitors. The fresh whole cell lysates were then addedto the ELISA plate for 2-hours. Between all subsequent steps plates werewashed 4 times with TBST. Detection antibody (recognizing thephospho-specific eptitope on GEF-H1b) was added for 1 hour, followed byaddition of an enzyme linked goat α-rabbit secondary antibody for 45minutes. Color development of the enzyme-linked antibody was performedwith a peroxidase substrate, ABTS (Moss, Inc.) with absorbance at 405 nMread with a spectrophotometer after 30 minute incubation. EC50 valueswere calculated by sigmoid curve fitting using a four-parameteranalysis.

PAK4 Kinase Domain Ki Data and PAK4 Cellular Assay EC50 Data of theCompounds of Examples 1-86:

Ki data EC50 Ex. # (uM) (nM) 1 0.041 >2000 2 0.014 36 3 0.011 16 4 0.115 0.22 87 6 0.0028 0.94 7 0.087 >4000 8 0.61 9 0.27 >4000 10 0.12 >400011 0.96 12 0.038 78 13 N/A >4000 14 0.27 15 0.090 500 16 0.076 1267 170.77 18 0.34 19 0.19 20 0.090 1199 21 0.24 22 0.72 23 0.14 >4000 24 0.6425 0.78 26 0.22 27 0.20 28 0.27 29 0.66 30 0.080 968 31 0.020 103 320.0035 19 33 0.047 618 34 0.021 778 35 0.26 262 36 >4000 37 0.27 805 380.096 1470 39 >4000 40 0.018 98 41 0.024 147 42 0.094 610 43 0.45 >400044 0.23 >4000 45 0.025 32 46 0.0072 21 47 0.37 401 48 1 49 0.16 332 500.016 340 51 0.018 31 52 0.0035 19 53 0.067 1745 54 0.14 55 >4000 560.24 570 57 0.94 58 0.014 2.5 59 0.17 2.4 60 0.040 <3.9 61 0.022 19 620.015 15 63 0.02 <3.9 64 0.003 <3.9 65 0.68 67.13 66 0.17 <3.9 67 0.2820. 68 0.0067 20 69 0.0019 3.0 70 0.0088 1.9 71 0.0052 72 0.016 4.4 730.0016 <3.9 74 0.0036 <3.9 75 0.0052 <3.9 76 0.0083 <3.9 77 0.093 69 780.33 >4000 79 0.039 <3.9 80 0.12 20 81 0.084 82 2.5 83 0.058 84 84 0.034455 85 0.10 86 0.055

We claim:
 1. A compound of formula I,

wherein: R¹ is chosen from —S(O)R^(a), —S(O)₂R^(a), C₁-C₁₂ alkyl, C₁-C₁₂alkyl substituted by 1 to 6 R⁵, C₃-C₁₂ cycloalkyl, C₃-C₁₂ cycloalkylsubstituted by 1 to 6 R⁵, C₂-C₁₂ alkenyl, C₂-C₁₂ alkenyl substituted by1 to 6 R⁵, C₄-C₁₂ cycloalkenyl, C₄-C₁₂ cycloalkenyl substituted by 1 to6 R⁵, C₂-C₁₂ alkynyl, C₂-C₁₂ alkynyl substituted by 1 to 6 R⁵, 3-12member heterocyclyl, 3-12 member heterocyclyl substituted by 1 to 6 R⁵,C₁-C₆ aralkyl, C₁-C₆ aralkyl substituted by 1 to 6 R⁵, C₁-C₆heteroaralkyl, C₁-C₆ heteroaralkyl substituted by 1 to 6 R⁵, C₆-C₁₀aryl, C₆-C₁₀ aryl substituted by 1 to 6 R⁵, 5-12 member heteroaryl, and5-12 member heteroaryl substituted by 1 to 6 R⁵, wherein any twoadjacent R⁵ together with the atoms to which they are attached may forma fused 4-7 member ring, and the said fused ring is optionally furthersubstituted by 1-3 R^(f); R² and R³ are each independently chosen from—H, C₁-C₆ perfluoroalkyl, C₁-C₆ alkyl, C₃-C₆ cycloalkyl, —(C₁-C₃alkylene)-(C₃-C₆ cycloalkyl), C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆alkoxy, -(L)_(m)-halide, -(L)_(m)-CN, -(L)_(m)-OH, -(L)_(m)-NH₂,-(L)_(m)-(C₁-C₆ monoalkylamino) and -(L)_(m)-(C₂-C₈ dialkylamino),provided that R² and R³ are not both H; or R² and R³ may form a ringselected from C₃-C₆ cycloalkyl, C₄-C₆ cycloalkenyl and 3-6 memberheterocyclyl, the said ring is optionally further substituted by 1 to 2groups selected from C₁-C₃ alkyl, C₁-C₃ perfluoroalkyl, C₁-C₃ alkoxy,oxo, —(C₁-C₃ alkylene)_(m)-halide, —(C₁-C₃ alkylene)_(m)-CN, —(C₁-C₃alkylene)_(m)-OH, —(C₁-C₃ alkylene)_(m)-NH₂, —(C₁-C₃alkylene)_(m)-(C₁-C₆ monoalkylamino) and —(C₁-C₃ alkylene)_(m)-(C₂-C₈dialkylamino); R⁴ is selected from R^(a), —C(O)R^(a), —C(O)NR^(a)R^(b),—C(O)OR^(a), —C(O)CH(R^(t))R^(a), —C(O)NHCH(R^(a))R^(b),—C(O)OCH(R^(a))R^(b), —C(O)CH(R^(t))CH(R^(a))R^(b), —C(O)SR^(a),—S(O)R^(a), —S(O)NR^(a)R^(b), —S(O)OR^(a), —S(O)₂R^(a),—S(O)₂NR^(a)R^(b) and —S(O)₂OR^(a), wherein R^(t) is H or C₁-C₃ alkyl;each R⁵ is independently selected from R^(c), -(L)_(m)-halide,-(L)_(m)-CN, -(L)_(m)-C(O)R^(c), -(L)_(m)-C(O)OR^(c),-(L)_(m)-C(O)NR^(c)R^(d), -(L)_(m)-C(O)SR^(c), -(L)_(m)-OR^(c),-(L)_(m)-OC(O)R^(c), -(L)_(m)-OC(O)NR^(c)R^(d), -(L)_(m)-O—C(O)OR^(c),-(L)_(m)-NO₂, -(L)_(m)-NR^(c)R^(d), -(L)_(m)-N(R^(c))C(O)R^(d),-(L)_(m)-N(R^(c))C(O)OR^(d), -(L)_(m)-NR^(c)S(O)R^(d),-(L)_(m)-NR^(c)S(O)OR^(d), -(L)_(m)-NR^(c)S(O)₂R^(d),-(L)_(m)-NR^(c)S(O)₂OR^(d), -(L)_(m)-SR^(c), -(L)_(m)-S(O)R^(c),-(L)_(m)-S(O)OR^(c), -(L)_(m)-S(O)₂R^(c), -(L)_(m)-S(O)₂OR^(c),-(L)_(m)-S(O)NR^(c)R^(d), -(L)_(m)-S(O)₂NR^(c)R^(d),-(L)_(m)-O-L-NR^(c)R^(d), -(L)_(m)-O-L-OR^(c) and-(L)_(m)-NR^(c)-L-OR^(d); each R^(a), R^(b), R^(c), and R^(d) isindependently selected from H, -(L)_(m)-(C₁-C₆ perfluoroalkyl), C₁-C₁₂alkyl, —(C₁-C₃ alkylene)_(m)-(C₃-C₁₂ cycloalkyl), —(C₃-C₅cycloalkylene)_(m)-(C₂-C₁₂ alkenyl), -(L)_(m)-(C₄-C₁₂ cycloakenyl),—(C₃-C₅ cycloalkylene)_(m)-(C₂-C₁₂ alkynyl), -(L)_(m)-(3-12 memberheterocyclyl), -(L)_(m)-(C₆-C₁₀ aryl) and -(L)_(m)-(5-12 memberheteroaryl), each R^(a), R^(b), R^(c) and R^(d) is independentlyoptionally further substituted by 1-6 R^(f); R^(a) and R^(b), or R^(c)and R^(d), together with the atom to which they are attached, mayoptionally form a ring selected from 3-12 member heterocyclyl and 5-12member heteroaryl, the said ring is optionally further substituted by1-6 R^(f); each R^(f) is independently selected from oxo, —(C₁-C₃alkylene)_(m)-(C₁-C₆ perfluoalkyl), C₁-C₁₂ alkyl, C₂-C₆ alkenyl, C₂-C₆alkynyl, —(C₁-C₃ alkylene)_(m)-(C₃-C₇ cycloalkyl), —(C₁-C₃alkylene)_(m)-(3-7 member heterocyclyl), —(C₁-C₃ alkylene)_(m)-(5-7member heteroaryl), -(L)_(m)-halide, -(L)_(m)-CN, -(L)_(m)-C(O)R^(k),-(L)_(m)-C(O)OR^(k), -(L)_(m)-C(O)NR^(k)R^(j), -(L)_(m)-OR^(k),-(L)_(m)-OC(O)R^(k), -(L)_(m)-NO₂, -(L)_(m)-NR^(k)R^(j),-(L)_(m)-N(R^(k))C(O)R^(j), -(L)_(m)-O-L-NR^(k)R^(j), -(L)_(m)-SR^(k),-(L)_(m)-S(O)R^(k), -(L)_(m)-S(O)₂R^(j)R^(k), each R^(f) isindependently optionally further substituted by 1-3 groups selected fromC₁-C₃ alkyl, halide and C₁-C₃ perfluoroalkyl; each R^(k) and R^(j) isindependently —H, —OH, C₁-C₃ perfluoroalkyl, C₁-C₆ alkyl, C₂-C₆ alkenyl,C₃-C₆ alkynyl, —(C₁-C₃ alkylene)_(m)-(C₃-C₆ cycloalkyl) or —(C₁-C₃alkylene)_(m)-(3 to 6 member heterocyclyl), R^(k) and R^(j) mayoptionally form a ring selected from 3-7 member heterocyclyl and 5-7member heteroaryl, the said ring is optionally further substituted by 1to 2 groups selected from C₁-C₃ alkyl, C₁-C₃ perfluoroalkyl, C₁-C₃alkoxy, oxo, —(C₁-C₃ alkylene)_(m)-halide, —(C₁-C₃ alkylene)_(m)-CN,—(C₁-C₃ alkylene)_(m)-OH, —(C₁-C₃ alkylene)_(m)-NH₂, —(C₁-C₃alkylene)_(m)-(C₁-C₆ monoalkylamino) and —(C₁-C₃ alkylene)_(m)-(C₂-C₈dialkylamino); each L is independently a bivalent radical selected from—(C₁-C₆ alkylene)-, —(C₃-C₇ cycloalkylene)-, —(C₁-C₆ alkylene)-(C₃-C₇cycloalkylene)- and —(C₃-C₇ cycloalkylene)-(C₁-C₆ alkylene)-; each m isindependently 0 or 1; and n is 1, 2, or 3; or a pharmaceuticallyacceptable salt thereof.
 2. The compound of claim 1, or apharmaceutically acceptable salt thereof, wherein n is 1; R² isunsubstituted methyl; R³ is unsubstituted methyl; R⁴ is—C(O)NHCH(R^(a))R^(b), —C(O)OCH(R^(a))R^(b) or—C(O)CH(R^(t))CH(R^(a))R^(b); R¹ is 5-12 member heteroaryl; and R¹ isoptionally further substituted as by 1-5 R⁵; each R⁵ is independently-(L¹)_(m)-(C₁-C₆ perfluoalkyl), C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆alkynyl, —(C₁-C₃ alkylene)_(m)-(C₃-C₄ cycloalkyl), —(C₁-C₃alkylene)_(m)-(3-4 member heterocyclyl) optionally substituted by 1-2C₁-C₃ alkyl, -(L¹)_(m)-halide, -(L¹)_(m)-CN, -(L¹)_(m)-C(O)R^(k),-(L¹)_(m)-C(O)OR^(k), -(L¹)_(m)-C(O)NR^(k)R^(j), -(L¹)_(m)-C(O)SR^(j),-(L¹)_(m)-OR^(k), -(L¹)_(m)-OC(O)R^(k), -(L¹)_(m)-OC(O)NR^(j)R^(k),-(L¹)_(m)-NO₂, -(L¹)_(m)-NR^(k)R^(j), -(L¹)_(m)-N(R^(k))C(O)R^(j),-(L¹)_(m)-N(R^(k))C(O)OR^(j), -(L¹)_(m)-O-L¹-NR^(k)R^(j),-(L¹)_(m)-O-L¹-OR^(k), -(L¹)_(m)-NR^(j)-L¹-OR^(k), -(L¹)_(m)-SR^(k),-(L¹)_(m)-S(O)R^(k), -(L¹)_(m)-S(O)OR^(k), -(L¹)_(m)-S(O)NR^(j)R^(k),-(L¹)_(m)-S(O)₂R^(k), -(L¹)_(m)-S(O)₂OR^(k) or-(L¹)_(m)-S(O)₂NR^(j)R^(k), wherein each R^(j); and R^(k) isindependently H, OH, C₁-C₃ alkyl or C₁-C₃ perfluoroalkyl, or R^(j); andR^(k) on the same nitrogen forms a 3-4 member ring selected fromaziridinyl and azetidinyl; L¹ is a bivalent radical selected from—(C₁-C₃ alkylene)-, —(C₃-C₄ cycloalkylene)-, -(3-4 memberheterocyclylene)-, —(C₁-C₃ alkylene)-(C₃-C₄ cycloalkylene)-, —(C₃-C₄cycloalkylene)-(C₁-C₃ alkylene)-, —(C₁-C₃ alkylene)-(3-4 memberheterocyclylene)- and -(3-4 member heterocyclylene)-(C₁-C₃ alkylene)-.3. A pharmaceutical composition comprising a compound of claim 1, or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable excipient.
 4. The compound of claim 1, or a pharmaceuticallyacceptable salt thereof, wherein n is
 1. 5. The compound of claim 1, ora pharmaceutically acceptable salt thereof, wherein each R² and R³ isindependently selected from H, unsubstituted C₁-C₃ alkyl andunsubstituted C₃-C₅ cycloalkyl, or R² and R³ form a ring selected fromunsubstituted cyclopropyl, unsubstituted cyclobutyl and unsubstitutedcyclopentyl.
 6. The compound of claim 1, or a pharmaceuticallyacceptable salt thereof, wherein R² is unsubstituted methyl and R³ isunsubstituted methyl.
 7. The compound of claim 1, or a pharmaceuticallyacceptable salt thereof, wherein R⁴ is selected from—C(O)NHCH(R^(a))R^(b), —C(O)OCH(R^(a))R^(b) andC(O)CH(R^(t))CH(R^(a))R^(b).
 8. The compound of claim 7, or apharmaceutically acceptable salt thereof, wherein R^(a) is selected fromphenyl, 5-12 member heteroaryl, 3-12 member heterocyclyl and 3-12 membercycloalkyl, where R^(a) is optionally further substituted by 1-6 R^(f),and R^(b) is a methyl group substituted by NR^(j)R^(k).